Pyrazolopyridine kinase inhibitors

ABSTRACT

The present invention relates to compounds of pyrazolopyridine derivatives useful as inhibitors of protein kinase. In one embodiment the compounds of the present invention are represented by the following structural formula: 
                         
or a pharmaceutically acceptable salt thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.61/298,649, filed Jan. 27, 2010 and U.S. Provisional Application No.61/410,426, filed Nov. 4, 2010, both of which are herein incorporated byreference.

BACKGROUND OF THE INVENTION

Protein kinases constitute a large family of structurally relatedenzymes that are responsible for the control of a variety of signaltransduction processes within the cell (see Hardie, G and Hanks, S. TheProtein Kinase Facts Book, I and II, Academic Press, San Diego, Calif.:1995).

In general, protein kinases mediate intracellular signaling by affectinga phosphoryl transfer from a nucleoside triphosphate to a proteinacceptor that is involved in a signaling pathway. These phosphorylationevents act as molecular on/off switches that can modulate or regulatethe target protein biological function. These phosphorylation events areultimately triggered in response to a variety of extracellular and otherstimuli. Examples of such stimuli include environmental and chemicalstress signals (e.g. shock, heat shock, ultraviolet radiation, bacterialendotoxin, and H₂O₂), cytokines (e.g. interleukin-1 (IL-1) and tumornecrosis factor alpha (TNF-a), and growth factors (e.g. granulocytemacrophage-colony stimulating factor (GM-CSF), and fibroblast growthfactor (FGF)). An extracellular stimulus may affect one or more cellularresponses related to cell growth, migration, differentiation, secretionof hormones, activation of transcription factors, muscle contraction,glucose metabolism, control of protein synthesis, survival andregulation of the cell cycle.

Kinases may be categorized into families by the substrates theyphosphorylate (e.g. protein-tyrosine, protein-serine/threonine, lipidsetc). Sequence motifs have been identified that generally correspond toeach of these kinase families (See, for example, Hanks, S. K., Hunter,T., FASEB J. 1995, 9, 576-596; Knighton et al., Science 1991, 253,407-414; Hiles et al, Cell 1992, 70, 419-429; Kunz et al, Cell 1993, 73,585-596; Garcia-Bustos et al, EMBO J 1994, 13, 2352-2361).

A serine/threonine kinase, protein kinase C-theta (PKC-theta), is amember of the novel, calcium independent PKC subfamily that isselectively expressed in T cells and skeletal muscle. Several lines ofevidence indicate that PKC-theta has an essential role in T cellactivation. Upon antigen stimulation of T cells, PKC-theta, but notother PKC isoforms, rapidly translocates from the cytoplasm to the siteof cell contact between the T cell and antigen-presenting cell (APC),where it localizes with the T cell receptor (TCR) in a region termed thecentral supramolecular activation cluster (cSMAC) (Monks et al., 1997,Nature, 385: 83-86; Monks et al., 1998, Nature, 395: 82-86).

It has been reported that PKC-theta selectively activates thetranscription factors AP-1 and NF-κB and integrates TCR and CD28co-stimulatory signals leading to the activation of the CD28 responseelement (CD28RE) in the IL-2 promoter (Baier-Bitterlich et al., 1996,Mol. Cell. Biol., 16: 1842-1850; Coudronniere et al., 2000, PNAS, 97:3394-3399). The specific role for PKC-theta in CD3/CD28 co-stimulationof T cells is highlighted in a study where expression of a kinase-deadPKC-theta mutant, or anti-sense PKC-theta dose-dependently inhibitedCD3/CD28 co-stimulated NF-κB activation, but not TNF-alpha-stimulatedNF-κB activation. This was not seen with other PKC isoforms (Lin et al.,2000, Mol. Cell. Biol., 20: 2933-2940). Recruitment of PKC-theta to theSMAC is reported to be mediated by its N-terminal regulatory domain andis necessary for T cell activation, as an over-expressed PKC-thetacatalytic fragment did not translocate and was unable to activate NF-κB,whereas a PKC-theta catalytic domain-Lck membrane-binding domain chimerawas able to reconstitute signaling (Bi et al., 2001, Nat. Immunol.,2:556-563).

Translocation of PKC-theta to the SMAC appears to be mediated by alargely PLC-gamma/DAG-independent mechanism, involving Vav andPI3-kinase (Villalba et al., 2002, JCB 157: 253-263), whilst activationof PKC-theta requires input from several signaling components includingLck, ZAP-70, SLP-76, PLC-gamma, Vav and PI3-kinase (Liu et al., 2000,JBC, 275: 3606-3609; Herndon et al., 2001, J. Immunol., 166: 5654-5664;Dienz et al., 2002, J. Immunol., 169: 365-372; Bauer et al., 2001 JBC.,276: 31627-31634). These biochemical studies in human T cells havegained credence from studies in PKC-theta knockout mice, which haveconfirmed a crucial role for this enzyme in T cell function.PKC-theta-/- mice are healthy and fertile, have a normally developedimmune system, but exhibit profound defects in mature T cell activation(Sun et al., 200, Nature, 404:402-407). Proliferative responses to TCRand TCR/CD28 co-stimulation were inhibited (>90%) as were in vivoresponses to antigen. In agreement with studies on human T cells,activation of the transcription factors AP-1 and NF-κB was abrogated,resulting in a severe deficit in IL-2 production and IL-2 R upregulation(Baier-Bitterlich et al., 1996, MBC, 16, 1842; Lin et al., 2000, MCB,20, 2933; Courdonniere, 2000, 97, 3394). More recently, studies inPKC-theta-deficient mice have indicated a role for PKC-theta in thedevelopment of mouse models of autoimmune diseases, including multiplesclerosis (MS), rheumatoid arthritis (RA) and irritable bowel disease(IBD) (Salek-Ardakani et al., 2006; Tan et al., 2006; Healy et al.,2006; Anderson et al., 2006). In these models, PKC-theta-deficient miceexhibited a marked reduction in disease severity that was associatedwith a profound defect in the development and effector function ofautoreactive T cells.

In addition to its role in T cell activation, PKC-theta is reported tomediate the phorbol ester-triggered survival signal that protects Tcells from Fas- and UV-induced apoptosis (Villalba et al., 2001, J.Immunol. 166: 5955-5963; Berttolotto et al., 2000, 275: 37246-37250).This pro-survival role is of interest because the human PKC-theta genehas been mapped to chromosome 10 (10p15), a region associated withmutations leading to T cell leukaemias and lymphomas (Erdel et al.,1995, Genomics 25: 295-297; Verma et al., 1987, J. Cancer Res. Clin.Oncol., 113: 192-196).

In vivo, the role for PKC-theta in immune responses to infection isdependent on the type of pathogen encountered. PKC-theta deficient miceelicit normal Th1 and cytotoxic T cell-mediated responses to severalviral infections and the protozoan parasite, Leishmania major andeffectively clear these infections (Marsland et al., 2004; Berg-Brown etal., 2004; Marsland et al., 2005; Giannoni et al., 2005). However,PKC-theta deficient mice are unable to wage normal Th2 T cell responsesagainst the parasite Nippostrongylus brasiliensis and certain allergens(Marsland et al., 2004; Salek-Ardakani et al., 2004) and are unable toclear Listeria monocytogenes infection (Sakowicz-Burkiewicz et al.,2008). Clearly in some circumstances, the requirement for PKC-theta in Tcell activation can be bypassed and this is likely to involve theprovision of additional signals to T cells, either from cells of theinnate immune system, or directly from the pathogen in the form ofpathogen associated molecular patterns (PAMPs) (Marsland et al., 2007).

More recently, studies in PKC-theta-deficient mice have indicated a rolefor PKC-theta in the development of mouse models of autoimmune diseases,including multiple sclerosis, rheumatoid arthritis and inflammatorybowel disease. In all cases where examined, PKC-theta-deficient miceexhibited a marked reduction in disease severity that was associatedwith a profound defect in the development of a newly discovered class ofT cells, Th17 cells (Salek-Ardakani et al., 2006; Tan et al., 2006;Healy et al., 2006; Anderson et al., 2006; Nagahama et al., 2008).PKC-theta therefore appears to be essential for the development ofpathogenic auto-reactive Th17 cells in the context of autoimmunity.These observations support the notion that targeting PKC-theta willprovide a way to target autoimmune T cell responses, leaving many T cellresponses (e.g., to viral infections) intact.

In addition to its role in T cell activation, PKC-theta mediates thephorbol ester-triggered survival signal that protects T cells from Fas-and UV-induced apoptosis (Villalba et al., 2001, J. Immunol. 166:5955-5963; Berttolotto et al., 2000, 275: 37246-37250). Thispro-survival role is of interest because the human PKC-theta gene hasbeen mapped to chromosome 10 (10p15), a region associated with mutationsleading to T cell leukaemias and lymphomas (Erdel et al., 1995, Genomics25: 295-297; Verma et al., 1987, J. Cancer Res. Clin. Oncol., 113:192-196).

Together, these data indicate that PKC-theta is an attractive target fortherapeutic intervention in inflammatory disorders, immune disorders,lymphomas and T cell leukaemias.

Accordingly, there is a great need to develop compounds useful asinhibitors of protein kinases. In particular, it would be desirable todevelop compounds that are useful as inhibitors of PKC-theta,particularly given the inadequate treatments currently available for themajority of the disorders implicated in their activation.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1, is an XRD of compound 9, as described herein.

SUMMARY OF THE INVENTION

This invention provides, in general, compounds that are useful as kinaseinhibitors. In one embodiment the compounds of the present invention arerepresented by structural formula I:

or a pharmaceutically acceptable salt thereof.

T is —NH— or absent.

Each J_(c1) and J_(c2) is independently —CN, —F, —Cl, —OR, —CH₂OR, or—CF₃.

Each U₁, U₂, and U₃ is independently —H, Z, or J_(b) wherein no morethan one of U₁, U₂, and U₃ is —H; or two of U₁, U₂, and U₃ join togetherto form a C1-C6 cyloalkyl ring having 0-1 heteroatoms independentlysubstituted with one or more J_(e).

Z is Y2-Q2.

Y2 is absent or C1-6 alkyl optionally and independently substituted withone or more J_(d).

Q2 is absent or C3-C8 cyloalkyl having 0-1 heteroatoms optionally andindependently substituted with one or more J_(e), wherein Y2 and Q2 arenot both absent (when U₁, U₂, and U₃ is Z).

Each J_(b) is independently —F, —OR, —CN, —CF₃, —N(R)₂, —C(O)N(R)₂, C1-6alkyl optionally and independently substituted with one or more J_(a).

Each J_(a) is independently —F, —OR, —N(R)₂, or —C(O)N(R)₂.

Each J_(d) is independently —OR, —CN, —C(O)N(R)₂, —N(R)₂ or F.

Each J_(e) is independently C1-C6 alkyl, —OR, —N(R)₂ CF₃, or F.

Each R is —H or C1-C6 alkyl.

Wherein there is a chiral center at the carbon indicated by *.

In one embodiment, the present invention is a method of treating orpreventing protein kinase-mediated condition in a subject, comprisingadministering to the subject an effective amount of a compound, apharmaceutically acceptable salt thereof, or composition of the presentinvention.

In one embodiment the present invention is the manufacture of acompound, a pharmaceutically acceptable salt thereof, or composition ofthe present invention for use in treating or preventing a proteinkinase-mediated condition in a subject.

In another embodiment, the compounds, pharmaceutically acceptable saltsthereof, and compositions of the present invention are also useful forthe study of kinases in biological and pathological phenomena; the studyof intracellular signal transduction pathways mediated by such kinases;and the comparative evaluation of new kinase inhibitors.

DETAILED DESCRIPTION OF THE INVENTION

This invention relates to compounds, pharmaceutically acceptable saltsthereof, and compositions (such as, pharmaceutical compositions) usefulas protein kinase inhibitors.

In one embodiment, the compounds, pharmaceutically acceptable saltsthereof, and compositions of the present invention are effective asinhibitors of PKCtheta.

Compounds of this invention include those described generally herein,and are further illustrated by the classes, subclasses, and speciesdisclosed herein. As used herein, the definitions defined herein shallapply unless otherwise indicated. For purposes of this invention, thechemical elements are identified in accordance with the Periodic Tableof the Elements, CAS version, Handbook of Chemistry and Physics, 75^(th)Ed. Additionally, general principles of organic chemistry are describedin “Organic Chemistry”, Thomas Sorrell, University Science Books,Sausalito: 1999, and “March's Advanced Organic Chemistry”, 5^(th) Ed.,Ed.: Smith, M. B. and March, J., John Wiley & Sons, New York: 2001, theentire contents of which are hereby incorporated by reference.

In one embodiment the compounds of the present invention are representedby structural formula I as described above.

In another embodiment, for compounds of formula I, U₁ is Z and U₃ isJ_(b) and the remainder of the variables are as described above.

In another embodiment, for compounds of formula I, U₁ and U₂ are Z andU₃ is J_(b) and the remainder of the variables are as described above.

In another embodiment, for compounds of formula I, Y2 is C1-C3 alkyloptionally and independently substituted with one or more J_(d). Q2 isabsent or C3-C6 alkyl optionally and independently substituted with oneor more J_(e). Each J_(d) is independently —OR, or F and the remainderof the variables are as described above.

In another embodiment, for compounds of formula I, J_(b) is —OH or —NH₂and the remainder of the variables are as described above.

In another embodiment, for compounds of formula I, J_(b) is —OH and theremainder of the variables are as described above.

In another embodiment, for compounds of formula I, each J_(c1) andJ_(c2) is independently —CF₃, —CN, —F, or —ClB and the remainder of thevariables are as described above.

In another embodiment, for compounds of formula I, each J_(c1) andJ_(c2) is independently —F, or —Cl and the remainder of the variablesare as described above.

In another embodiment, for compounds of formula I, each J_(c1) andJ_(c2) is —F and the remainder of the variables are as described above.

In another embodiment, for compounds of formula I, J_(c1) is F andJ_(c1) is Cl; or J_(c1) is Cl and J_(c2) is F.

In certain embodiments the compound of formula I is not:

In one embodiment the compounds of the present invention are representedby structural formula II

or a pharmaceutically acceptable salt thereof, wherein:

T is —CH₂—, —CH(J_(b))-, —C(J_(b))₂-, —NH— or —N(J_(b))-.

t is 0, 1, or 2.

w is 0 or 1.

Each J, is independently CN, F, Cl, —OR, —CH₂OR, or CF₃.

U is Z or J_(b).

Z is Y2-Q2.

Y2 is absent or C1-6 alkyl optionally and independently substituted withone or more J_(d).

Q2 is absent or C3-C8 cyloalkyl having 0-1 heteroatoms optionally andindependently substituted with one or more J_(e); wherein Y2 and Q2 arenot both absent.

Each J_(b) is independently —F, —OR, —CN, —CF₃, —N(R)₂, —C(O)N(R)₂, C1-6alkyl optionally and independently substituted with one or more J_(a).

Each J_(a) is independently —F, —OR, —N(R)₂, or —C(O)N(R)₂.

Each J_(d) is independently —OR, —CN, —C(O)N(R)₂, —N(R)₂ or F.

Each J_(e) is independently —OR, CF₃, —N(R)₂ or F.

Each R is —H or C1-C6 alkyl.

With the proviso that the compound is not:

In one embodiment the compound of structural formula II has thefollowing structural formula:

As described herein, a specified number range of atoms includes anyinteger therein. For example, a group having from 1-4 atoms could have1, 2, 3, or 4 atoms.

As used here the terms “absent” and “a bond” can be used interchangeablyto mean the variable does not exits in that embodiment, that is thevariable does not represent an atom or groups of atoms.

The term “stable”, as used herein, refers to compounds that are notsubstantially altered when subjected to conditions to allow for theirproduction, detection, recovery, storage, purification, and use for oneor more of the purposes disclosed herein. In some embodiments, a stablecompound or chemically feasible compound is one that is notsubstantially altered when kept at a temperature of 40° C. or less, inthe absence of moisture or other chemically reactive conditions, for atleast a week.

The term “aliphatic” or “aliphatic group”, as used herein, means astraight-chain (i.e., unbranched), or branched, hydrocarbon chain thatis completely saturated or that contains one or more units ofunsaturation but is non-aromatic.

Unless otherwise specified, aliphatic groups contain 1-20 aliphaticcarbon atoms. In some embodiments, aliphatic groups contain 1-10aliphatic carbon atoms. In other embodiments, aliphatic groups contain1-8 aliphatic carbon atoms. In still other embodiments, aliphatic groupscontain 1-6 aliphatic carbon atoms, and in yet other embodimentsaliphatic groups contain 1-4 aliphatic carbon atoms. Aliphatic groupsmay be linear or branched alkyl, alkenyl, or alkynyl groups. Specificexamples include, but are not limited to, methyl, ethyl, isopropyl,n-propyl, sec-butyl, vinyl, n-butanol, thinly, and tert-butyl.

The term “alkyl” as used herein means a saturated straight or branchedchain hydrocarbon. The term “alkenyl” as used herein means a straight orbranched chain hydrocarbon comprising one or more double bonds. The term“alkynyl” as used herein means a straight or branched chain hydrocarboncomprising one or more triple bonds.

The term “cycloaliphatic” (or “carbocyclic” or “carbocyclic” or“carbocyclic”) refers to a non-aromatic monocyclic carbon containingring which can be saturated or contain one or more units ofunsaturation, having three to fourteen ring carbon atoms. The termincludes polycyclic fused, spiro or bridged carbocyclic ring systems.The term also includes polycyclic ring systems in which the carbocyclicring can be fused to one or more non-aromatic carbocyclic orheterocyclic rings or one or more aromatic rings or combination thereof,wherein the radical or point of attachment is on the carbocyclic ring.Fused bicyclic ring systems comprise two rings which share two adjoiningring atoms, bridged bicyclic group comprise two rings which share threeor four adjacent ring atoms, spiro bicyclic ring systems share one ringatom. Examples of cycloaliphatic groups include, but are not limited to,cycloalkyl and cycloalkenyl groups. Specific examples include, but arenot limited to, cyclohexyl, cyclopropentyl, cyclopropyl and cyclobutyl.

The term “heterocycle” (or “heterocyclyl”, or “heterocyclic”) as usedherein means refers to a non-aromatic monocyclic ring which can besaturated or contain one or more units of unsaturation, having three tofourteen ring atoms in which one or more ring carbons is replaced by aheteroatom such as, N, S, or O. The term includes polycyclic fused,spiro or bridged heterocyclic ring systems. The term also includespolycyclic ring systems in which the heterocyclic ring can be fused toone or more non-aromatic carbocyclic or heterocyclic rings or one ormore aromatic rings or combination thereof, wherein the radical or pointof attachment is on the heterocyclic ring. Examples of heterocyclesinclude, but are not limited to, piperidinyl, piperizinyl, pyrrolidinyl,pyrazolidinyl, imidazolidinyl, azepanyl, diazepanyl, triazepanyl,azocanyl, diazocanyl, triazocanyl, oxazolidinyl, isoxazolidinyl,thiazolidinyl, isothiazolidinyl, oxazocanyl, oxazepanyl, thiazepanyl,thiazocanyl, benzimidazolonyl, tetrahydrofuranyl, tetrahydrothiophenyl,tetrahydrothiophenyl, morpholino, including, for example, 3-morpholino,4-morpholino, 2-thiomorpholino, 3-thiomorpholino, 4-thiomorpholino,1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, 1-piperazinyl,2-piperazinyl, 3-piperazinyl, 1-pyrazolinyl, 3-pyrazolinyl,4-pyrazolinyl, 5-pyrazolinyl, 1-piperidinyl, 2-piperidinyl,3-piperidinyl, 4-piperidinyl, 2-thiazolidinyl, 3-thiazolidinyl,4-thiazolidinyl, 1-imidazolidinyl, 2-imidazolidinyl, 4-imidazolidinyl,5-imidazolidinyl, indolinyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, benzothiolanyl, benzodithianyl,dihydro-benzimidazol-2-onyl, and 1,3-dihydro-imidazol-2-onyl,azabicyclopentyl, azabicyclohexyl, azabicycloheptyl, azabicyclooctyl,azabicyclononyl, azabicyclodecyl, diazabicyclohexyl, diazabicycloheptyl,dihydroindazolyl, dihydrobenzimidazolyl, tetrahydropyridyl,dihydropyridyl, tetrahydropyrazinyl, dihydropyrazinyl,tetrahydropyrimidinyl, dihydropyrimidinyl, dihydropyrrolyl,dihydropyrazolyl, dihydroimidazolyl, octahydropyrrolopyrazyl,octahydropyrrolopyridyl, octahydropyridopyrazyl, octahydropyridopyridyl,diazabicyclooctyl, diazabicyclononyl, and diazabicyclodecyl.

As used herein, unless stated otherwise, bicyclic rings can be fused,spiro and bridged.

The term “heteroatom” means one or more of oxygen, sulfur, nitrogen,phosphorus, or silicon (including, any oxidized form of nitrogen,sulfur, phosphorus, or silicon; the quaternized form of any basicnitrogen or; a substitutable nitrogen of a heterocyclic ring, forexample N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) orNR⁺ (as in N-substituted pyrrolidinyl)).

The term “unsaturated”, as used herein, means that a moiety has one ormore units of unsaturation.

The term “alkoxy”, or “thioalkyl”, as used herein, refers to an alkylgroup, as previously defined, attached to the molecule through an oxygen(“alkoxy” e.g., —O-alkyl) or sulfur (“thioalkyl” e.g., —S-alkyl) atom.

The terms “haloalkyl”, “haloalkenyl”, “halo aliphatic”, and “haloalkoxy”(or “aminoalkyl”, “hydroxyalkyl” etc.,) mean alkyl, alkenyl or alkoxy,as the case may be, substituted with one or more halogen atoms. Thisterm includes perfluorinated alkyl groups, such as —CF₃ and —CF₂CF₃.

The terms “halogen”, “halo”, and “hal” mean F, Cl, Br, or I. The termhalo aliphatic and —O(halo aliphatic) include, mono- di- and tri- halosubstituted aliphatic groups.

The term “aryl” used alone or as part of a larger moiety as in “aalkyl”, “aralkoxy”, “aryloxyalkyl”, or “heteroaryl” refers tocarbocyclic and or heterocyclic aromatic ring systems. The term “aryl”may be used interchangeably with the term “aryl ring”.

Carbocyclic aromatic ring groups have only carbon ring atoms (typicallysix to fourteen) and include monocyclic aromatic rings such as phenyland fused polycyclic aromatic ring systems in which two or morecarbocyclic aromatic rings are fused to one another. Examples include1-naphthyl, 2-naphthyl, 1-anthracyl and 2-anthracyl. Also includedwithin the scope of the term “carbocyclic aromatic ring”, as it is usedherein, is a group in which an aromatic ring is fused to one or morenon-aromatic rings (carbocyclic or heterocyclic), such as in an indanyl,phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl,where the radical or point of attachment is on the aromatic ring.

The term “heteroaryl”, “heteroaromatic”, “heteroaryl ring”, “heteroarylgroup” and “heteroaromatic group”, used alone or as part of a largermoiety as in “heteroaralkyl” or “heteroarylalkoxy”, refers toheteroaromatic ring groups having five to fourteen members, includingmonocyclic heteroaromatic rings and polycyclic aromatic rings in which amonocyclic aromatic ring is fused to one or more other aromatic ring.Heteroaryl groups have one or more ring heteroatoms. Also includedwithin the scope of the term “heteroaryl”, as it is used herein, is agroup in which an aromatic ring is fused to one or more non-aromaticrings (carbocyclic or heterocyclic), where the radical or point ofattachment is on the aromatic ring. Bicyclic 6,5 heteroaromatic ring, asused herein, for example, is a six membered heteroaromatic ring fused toa second five membered ring, wherein the radical or point of attachmentis on the six membered ring. Examples of heteroaryl groups includepyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, imidazolyl, pyrrolyl,pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl,thiazolyl, isothiazolyl or thiadiazolyl including, for example,2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl,5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxadiazolyl,5-oxadiazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 3-pyrazolyl,4-pyrazolyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl,4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 3-pyridazinyl,2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-triazolyl, 5-triazolyl,tetrazolyl, 2-thienyl, 3-thienyl, carbazolyl, benzimidazolyl,benzothienyl, benzofuranyl, indolyl, benzotriazolyl, benzothiazolyl,benzoxazolyl, benzimidazolyl, isoquinolinyl, indolyl, isoindolyl,acridinyl, benzisoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl,1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl,1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, purinyl,pyrazinyl, 1,3,5-triazinyl, quinolinyl (e.g., 2-quinolinyl,3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g., 1-isoquinolinyl,3-isoquinolinyl, or 4-isoquinolinyl).

The term “protecting group” and “protective group” as used herein, areinterchangeable and refer to an agent used to temporarily block one ormore desired functional groups in a compound with multiple reactivesites. In certain embodiments, a protecting group has one or more, orpreferably all, of the following characteristics: a) is addedselectively to a functional group in good yield to give a protectedsubstrate that is b) stable to reactions occurring at one or more of theother reactive sites; and c) is selectively removable in good yield byreagents that do not attack the regenerated, deprotected functionalgroup. As would be understood by one skilled in the art, in some cases,the reagents do not attack other reactive groups in the compound. Inother cases, the reagents may also react with other reactive groups inthe compound. Examples of protecting groups are detailed in Greene, T.W., Wuts, P. G in “Protective Groups in Organic Synthesis”, ThirdEdition, John Wiley & Sons, New York: 1999 (and other editions of thebook), the entire contents of which are hereby incorporated byreference. The term “nitrogen protecting group”, as used herein, refersto an agent used to temporarily block one or more desired nitrogenreactive sites in a multifunctional compound. Preferred nitrogenprotecting groups also possess the characteristics exemplified for aprotecting group above, and certain exemplary nitrogen protecting groupsare also detailed in Chapter 7 in Greene, T. W., Wuts, P. G in“Protective Groups in Organic Synthesis”, Third Edition, John Wiley &Sons, New York: 1999, the entire contents of which are herebyincorporated by reference.

In some embodiments, where indicated a methylene unit of an aliphaticchain is optionally replaced with another atom or group. Examples ofsuch atoms or groups include, but are not limited to, G which includes,—N(R′)—, —O—, —C(O)—, —C(═N—CN)—, —C(═NR′)— —C(═NOR′)—, —S—, —S(O)—, and—S(O)₂—. These atoms or groups can be combined to form larger groups.Examples of such larger groups include, but are not limited to, —OC(O)—,—C(O)CO—, —CO₂—, —C(O)NR′′—, —C(═N—CN), —N(R′)C(O)—, —N(R′)C(O)O—,—S(O)₂N(R′)—, —N(R)SO₂—, —N(R′)C(O)N(R′)—, —OC(O)N(R′)—, and—N(R)SO₂N(R′)—, wherein R′ is defined herein.

Only those replacement and combinations of groups that result in astable structure are contemplated. Optional replacements can occur bothwithin the chain and/or at either end of the chain; i.e. both at thepoint of attachment and/or also at the terminal end. Two optionalreplacements can also be adjacent to each other within a chain so longas it results in a chemically stable compound.

In some embodiments the optional replacements can also completelyreplace all of the carbon atoms in a chain. For example, a C₃ aliphaticcan be optionally replaced by —N(R′)—, —C(O)—, and —N(R′)— to form—N(R′)C(O)N(R′)—(a urea), or a C₁ aliphatic can be optionally bereplaced by, for example, —O—, NH— etc. In certain instances of theseembodiments the chain is a linker.

Unless otherwise indicated, if the replacement occurs at the terminalend, the replacement atom is bound to an H on the terminal end. Forexample, if —CH₂CH₂CH₃ were optionally replaced with —O—, the resultingcompound could be —OCH₂CH₃, —CH₂OCH₃, or —CH₂CH₂OH, or if —CH₂CH₃ wereoptionally replaced with —O—, the resulting compound could be —OCH₃, or—CH₂CH₂OH, or if —CH₂CH₃ were optionally replaced with —C(O)—, theresulting compound could be —C(O)CH₃, or —CH₂C(O)H.

In an alternative embodiment where specified herein, aliphatic chains inwhich up to three (0-3) methylene groups are optionally replaced by G′,wherein G′ is —N(R′)—, —O—, —C(O)—, or —S(O)_(p)—, (wherein R′ and p areas defined herein) require at least one unreplaced methylene group(—CH(substituent)- or —CH₂—) in the chain. For example, the methylenegroup in a C₁ aliphatic cannot be replaced by, for example, —OH, —NH₂etc., to give —OH and —NH₂ as the substituent without any methylenegroup in the chain, or ii) two methylene groups in a C₂ aliphatic groupscannot be replaced by —C(O)— and —O— to give —C(O)OH. In certaininstances of these alternative embodiment the chain is not a linker butrather a substituent only joined to the rest of the molecule in oneplace. These aliphatic groups are further substituted as defined herein.

Unless otherwise indicated, structures depicted herein are also meant toinclude all isomeric (e.g., enantiomeric, diastereomeric, geometric,conformational, and rotational) forms of the structure. For example, the(R) and (S) configurations for each asymmetric center, (Z) and (E)double bond isomers, and (Z) and (E) conformational isomers are includedin this invention. The use of (R) and (S) in the structures hereinrepresents stereochemistry and is distinct from the atom sulphur S orthe variable R in the description of the variables. As would beunderstood to one skilled in the art, a substituent can freely rotatearound any rotatable bonds. For example, a substituent drawn as

also represents

Therefore, single stereochemical isomers as well as enantiomeric,diastereomeric, geometric, conformational, and rotational mixtures ofthe present compounds are within the scope of the invention.

The preferred conformation of the compounds of the invention is (S)where indicated in the piperazine ring and (R) on the alpha carbon.

The stereochemistry of the instant molecules can be determined usingX-ray diffraction, using techniques know to those of skill in the art.In an alternative embodiment, without wishing to be bound by theory, thestereochemistry can be predicted based on the PKCtheta potency in theassays described herein.

Unless otherwise indicated, all tautomeric forms of the compounds of theinvention are within the scope of the invention.

Additionally, unless otherwise indicated, structures depicted herein arealso meant to include compounds that differ only in the presence of oneor more isotopically enriched atoms. For example, compounds having thepresent structures except for the replacement of hydrogen by deuteriumor tritium, or the replacement of a carbon by a ¹³C- or ¹⁴C-enrichedcarbon are within the scope of this invention. Such compounds areuseful, for example, as analytical tools or probes in biological assays.

As described herein, where indicated compounds of the invention mayoptionally be substituted with one or more substituents, such as areillustrated generally herein, or as exemplified by particular classes,subclasses, and species of the invention. It will be appreciated thatthe phrase “optionally substituted” is used interchangeably with thephrase “substituted or unsubstituted.” In general, the term“substituted”, whether preceded by the term “optionally” or not, refersto the replacement of hydrogen radicals in a given structure with theradical of a specified substituent. Unless otherwise indicated, anoptionally substituted group may have a substituent at eachsubstitutable position of the group, and when more than one position inany given structure may be substituted with more than one substituentselected from a specified group, the substituent may be either the sameor different at every position.

Only those choices and combinations of substituents that result in astable structure are contemplated. Such choices and combinations will beapparent to those of ordinary skill in the art and may be determinedwithout undue experimentation.

The term “ring atom” is an atom such as C, N, O or S that is in the ringof an aromatic group, cycloalkyl group or non-aromatic heterocyclicring.

A “substitutable ring atom” in an aromatic or non-aromatic ring group isa ring carbon or nitrogen atom bonded to a hydrogen atom. The hydrogencan be optionally replaced with a suitable substituent group. Thus, theterm “substitutable ring atom” does not include ring nitrogen or carbonatoms which are shared when two rings are fused. In addition,“substitutable ring atom” does not include ring carbon or nitrogen atomswhen the structure depicts that they are already attached to a moietyother than hydrogen.

An optionally substituted aryl group as defined herein may contain oneor more substitutable ring atoms, which may be bonded to a suitablesubstituent. Examples of suitable substituents on a substitutable ringcarbon atom of an aryl group includes R@. R@ is —Ra, —Br, —Cl, —I, —F,—ORa, —SRa, —O—CORa, —CORa, —CSRa, —CN, —NO₂, —NCS, —SO₃H, —N(RaRb),—COORa, —NRcNRcCORa, —NRcNRcCO₂Ra, —CHO, —CON(RaRb), —OC(O)N(RaRb),—CSN(RaRb), —NRcCORa, —NRcCOORa, —NRcCSRa, —NRcCON(RaRb),—NRcNRcC(O)N(RaRb), —NRcCSN(RaRb), —C(═NRc)-N(RaRb), —C(═S)N(RaRb),—NRd-C(═NRc)-N(RaRb), —NRcNRaRb, —S(O)_(p)NRaRb, —NRcSO₂N(RaRb),—NRcS(O)_(p)Ra, —S(O)_(p)Ra, —OS(O)_(p)NRaRb or —OS(O)_(p)Ra; wherein pis 1 or 2.

Ra-Rd are each independently —H, an aliphatic group, aromatic group,non-aromatic carbocyclic or heterocyclic group or —N(RaRb), takentogether, form a non-aromatic heterocyclic group. The aliphatic,aromatic and non-aromatic heterocyclic group represented by Ra-Rd andthe non-aromatic heterocyclic group represented by —N(RaRb) are eachoptionally and independently substituted with one or more groupsrepresented by R^(#). Preferably Ra-Rd are unsubstituted.

R^(#) is halogen, R⁺, —OR⁺, —SR⁺, —NO₂, —CN, —N(R⁺)₂, —COR⁺, —COOR⁺,—NHCO₂R⁺, —NHC(O)R⁺, —NHNHC(O)R⁺, —NHC(O)N(R⁺)₂, —NHNHC(O)N(R⁺)₂,—NHNHCO₂R⁺, —C(O)N(R⁺)₂, —OC(O)R⁺, —OC(O)N(R⁺)₂, —S(O)₂R⁺, —SO₂N(R⁺)₂,—S(O)R⁺, —NHSO₂N(R⁺)₂, —NHSO₂R⁺, —C(═S)N(R⁺)₂, or —C(═NH)—N(R⁺)₂.

R⁺ is —H, a C1-C4 alkyl group, a monocyclic aryl group, a non-aromaticcarbocyclic or heterocyclic group each optionally substituted withalkyl, haloalkyl, alkoxy, haloalkoxy, halogen, —CN, —NO₂, amine,alkylamine or dialkylamine Preferably R+ is unsubstituted.

An optionally substituted aliphatic or a non-aromatic heterocyclic orcarbocyclic group as used herein may contain one or more substituents.Examples of suitable substituents for an aliphatic group or a ringcarbon of a non-aromatic heterocyclic group is R″. R″ includes thosesubstituents listed above for R@ and ═O, ═S, ═NNHR**, ═NN(R**)2,═NNHC(O)R**, ═NNHCO2 (alkyl), ═NNHSO2 (alkyl), ═NR**, spiro cycloalkylgroup or fused cycloalkyl group. Each R** is independently selected fromhydrogen, an unsubstituted alkyl group or a substituted alkyl group.Examples of substituents on the alkyl group represented by R** includeamino, alkylamino, dialkylamino, aminocarbonyl, halogen, alkyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylaminocarbonyloxy,dialkylaminocarbonyloxy, alkoxy, nitro, cyano, carboxy, alkoxycarbonyl,alkylcarbonyl, hydroxy, haloalkoxy, or haloalkyl.

When a heterocyclyl, heteroaryl, or heteroaralkyl group contains anitrogen atom, it may be substituted or unsubstituted. When a nitrogenatom in the aromatic ring of a heteroaryl group has a substituent thenitrogen may be a quaternary nitrogen.

A preferred position for substitution of a non-aromaticnitrogen-containing heterocyclic group is the nitrogen ring atom.Suitable substituents on the nitrogen of a non-aromatic heterocyclicgroup or heteroaryl group include —R^, —N(R^)₂, C(O)R^, CO₂R^,—C(O)C(O)R^, —SO₂R^, SO₂N(R^)₂, C(═S)N(R^)₂, C(═NH)—N(R^)₂, and—NR^SO₂R^; wherein RA is hydrogen, an aliphatic group, a substitutedaliphatic group, aryl, substituted aryl, heterocyclic or carbocyclicring or a substituted heterocyclic or carbocyclic ring. Examples ofsubstituents on the group represented by R′′ include alkyl, haloalkoxy,haloalkyl, alkoxyalkyl, sulfonyl, alkylsulfonyl, halogen, nitro, cyano,hydroxy, aryl, carbocyclic or heterocyclic ring, oxo, amino, alkylamino,dialkylamino, aminocarbonyl, alkylaminocarbonyl,dialkylaminocarbonyloxy, alkoxy, carboxy, alkoxycarbonyl, oralkylcarbonyl. Preferably R′′ is not substituted.

Non-aromatic nitrogen containing heterocyclic rings that are substitutedon a ring nitrogen and attached to the remainder of the molecule at aring carbon atom are said to be N substituted. For example, an N alkylpiperidinyl group is attached to the remainder of the molecule at thetwo, three or four position of the piperidinyl ring and substituted atthe ring nitrogen with an alkyl group. Non-aromatic nitrogen containingheterocyclic rings such as pyrazinyl that are substituted on a ringnitrogen and attached to the remainder of the molecule at a second ringnitrogen atom are said to be N′ substituted-N-heterocycles. For example,an N′ acyl N-pyrazinyl group is attached to the remainder of themolecule at one ring nitrogen atom and substituted at the second ringnitrogen atom with an acyl group.

As used herein an optionally substituted aralkyl can be substituted onboth the alkyl and the aryl portion. Unless otherwise indicated as usedherein optionally substituted aralkyl is optionally substituted on thearyl portion.

The compounds of the invention are defined herein by their chemicalstructures and/or chemical names. Where a compound is referred to byboth a chemical structure and a chemical name, and the chemicalstructure and chemical name conflict, the chemical structure isdeterminative of the compound's identity.

The compounds of this invention can exist in free form for treatment, orwhere appropriate, as a pharmaceutically acceptable salt.

As used herein, the term “pharmaceutically acceptable salt” refers tosalts of a compound which are, within the scope of sound medicaljudgment, suitable for use in contact with the tissues of humans andlower animals without undue side effects, such as, toxicity, irritation,allergic response and the like, and are commensurate with a reasonablebenefit/risk ratio.

Pharmaceutically acceptable salts are well known in the art. Forexample, S. M. Berge et al., describe pharmaceutically acceptable saltsin detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporatedherein by reference. Pharmaceutically acceptable salts of the compoundsof this invention include those derived from suitable inorganic andorganic acids and bases. These salts can be prepared in situ during thefinal isolation and purification of the compounds. Acid addition saltscan be prepared by 1) reacting the purified compound in its free-basedform with a suitable organic or inorganic acid and 2) isolating the saltthus formed.

Examples of pharmaceutically acceptable, nontoxic acid addition saltsare salts of an amino group formed with inorganic acids such ashydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid andperchloric acid or with organic acids such as acetic acid, oxalic acid,maleic acid, tartaric acid, citric acid, succinic acid or malonic acidor by using other methods used in the art such as ion exchange. Otherpharmaceutically acceptable salts include adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, glycolate, gluconate, glycolate,hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate,lauryl sulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, salicylate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like.

Base addition salts can be prepared by 1) reacting the purified compoundin its acid form with a suitable organic or inorganic base and 2)isolating the salt thus formed. Salts derived from appropriate basesinclude alkali metal (e.g., sodium, lithium, and potassium), alkalineearth metal (e.g., magnesium and calcium), ammonium and N⁺ (C₁₋₄alkyl)₄salts. This invention also envisions the quaternization of any basicnitrogen-containing groups of the compounds disclosed herein. Water oroil-soluble or dispersible products may be obtained by suchquaternization.

Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, loweralkyl sulfonate and aryl sulfonate. Other acids and bases,while not in themselves pharmaceutically acceptable, may be employed inthe preparation of salts useful as intermediates in obtaining thecompounds of the invention and their pharmaceutically acceptable acid orbase addition salts.

It should be understood that this invention includesmixtures/combinations of different pharmaceutically acceptable salts andalso mixtures/combinations of compounds in free form andpharmaceutically acceptable salts.

In addition to the compounds of this invention, pharmaceuticallyacceptable solvates (e.g., hydrates) and clathrates of the compounds ofthis invention may also be employed in compositions to treat or preventthe herein identified disorders.

As used herein, the term “pharmaceutically acceptable solvate,” is asolvate formed from the association of one or more pharmaceuticallyacceptable solvent molecules to one of the compounds the invention. Theterm solvate includes hydrates (e.g., hemihydrate, monohydrate,dihydrate, trihydrate, tetrahydrate, and the like).

As used herein, the term “hydrate” means a compound of the presentinvention or a salt thereof, that further includes a stoichiometric ornon-stoichiometric amount of water bound by non-covalent intermolecularforces.

As used herein, the term “clathrate” means a compound of the presentinvention or a salt thereof in the form of a crystal lattice thatcontains spaces (e.g., channels) that have a guest molecule (e.g., asolvent or water) trapped within.

In addition to the compounds of this invention, pharmaceuticallyacceptable derivatives or prodrugs, and esters, of the compounds of thisinvention may also be employed in compositions to treat or prevent theherein identified disorders.

As used herein and unless otherwise indicated, the term “prodrug” meansa derivative of a compound that can hydrolyze, oxidize, or otherwisereact under biological conditions (in vitro or in vivo) to provide acompound of this invention. Prodrugs may become active upon suchreaction under biological conditions, or they may have activity in theirunreacted forms. Examples of prodrugs contemplated in this inventioninclude, but are not limited to, analogs or derivatives of compounds ofthe invention that comprise biohydrolyzable moieties such asbiohydrolyzable amides, biohydrolyzable esters, biohydrolyzablecarbamates, biohydrolyzable carbonates, biohydrolyzable ureides, andbiohydrolyzable phosphate analogues. Other examples of prodrugs includederivatives of compounds of the invention that comprise —NO, —NO2, —ONO,or —ONO2 moieties. Prodrugs can typically be prepared using well-knownmethods, such as those described by BURGER'S MEDICINAL CHEMISTRY ANDDRUG DISCOVERY (1995) 172-178, 949-982 (Manfred E. Wolff ed., 5th ed).

A “pharmaceutically acceptable derivative” is an adduct or derivativewhich, upon administration to a patient in need, is capable ofproviding, directly or indirectly, a compound as otherwise describedherein, or a metabolite or residue thereof. Examples of pharmaceuticallyacceptable derivatives include, but are not limited to, esters and saltsof such esters.

A “pharmaceutically acceptable derivative or prodrug” includes anypharmaceutically acceptable ester, salt of an ester or other derivativeor salt thereof of a compound, of this invention which, uponadministration to a recipient, is capable of providing, either directlyor indirectly, a compound of this invention or an inhibitorily activemetabolite or residue thereof. Particularly favoured derivatives orprodrugs are those that increase the bioavailability of the compounds ofthis invention when such compounds are administered to a patient (e.g.,by allowing an orally administered compound to be more readily absorbedinto the blood) or which enhance delivery of the parent compound to abiological compartment (e.g., the brain or lymphatic system) relative tothe parent species.

Pharmaceutically acceptable prodrugs of the compounds of this inventioninclude, without limitation, esters, amino acid esters, phosphateesters, metal salts and sulfonate esters.

As used herein, the phrase “side effects” encompasses unwanted andadverse effects of a therapy (e.g., a prophylactic or therapeuticagent). Side effects are always unwanted, but unwanted effects are notnecessarily adverse. An adverse effect from a therapy (e.g.,prophylactic or therapeutic agent) might be harmful or uncomfortable orrisky. Side effects include, but are not limited to fever, chills,lethargy, gastrointestinal toxicities (including gastric and intestinalulcerations and erosions), nausea, vomiting, neurotoxicities,nephrotoxicities, renal toxicities (including such conditions aspapillary necrosis and chronic interstitial nephritis), hepatictoxicities (including elevated serum liver enzyme levels),myelotoxicities (including leukopenia, myelosuppression,thrombocytopenia and anemia), dry mouth, metallic taste, prolongation ofgestation, weakness, somnolence, pain (including muscle pain, bone painand headache), hair loss, asthenia, dizziness, extra-pyramidal symptoms,akathisia, cardiovascular disturbances and sexual dysfunction.

In one embodiment the present invention is a pharmaceutical compositioncomprising a compound of the present invention and a pharmaceuticallyacceptable carrier, diluent, adjuvant or vehicle. In one embodiment thepresent invention is a pharmaceutical composition comprising aneffective amount of compound of the present invention and apharmaceutically acceptable carrier, diluent, adjuvant or vehicle.Pharmaceutically acceptable carriers include, for example,pharmaceutical diluents, excipients or carriers suitably selected withrespect to the intended form of administration, and consistent withconventional pharmaceutical practices.

A pharmaceutically acceptable carrier may contain inert ingredientswhich do not unduly inhibit the biological activity of the compounds.The pharmaceutically acceptable carriers should be biocompatible, e.g.,non-toxic, non-inflammatory, non-immunogenic or devoid of otherundesired reactions or side-effects upon the administration to asubject. Standard pharmaceutical formulation techniques can be employed.

The pharmaceutically acceptable carrier, adjuvant, or vehicle, as usedherein, includes any and all solvents, diluents, or other liquidvehicle, dispersion or suspension aids, surface active agents, isotonicagents, thickening or emulsifying agents, preservatives, solid binders,lubricants and the like, as suited to the particular dosage formdesired. Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W.Martin (Mack Publishing Co., Easton, Pa., 1980) discloses variouscarriers used in formulating pharmaceutically acceptable compositionsand known techniques for the preparation thereof. Except insofar as anyconventional carrier medium is incompatible with the compounds of theinvention, such as by producing any undesirable biological effect orotherwise interacting in a deleterious manner with any othercomponent(s) of the pharmaceutically acceptable composition, its use iscontemplated to be within the scope of this invention.

Some examples of materials which can serve as pharmaceuticallyacceptable carriers include, but are not limited to, ion exchangers,alumina, aluminum stearate, lecithin, serum proteins, such as humanserum albumin, buffer substances such as phosphates, glycine, sorbicacid, or potassium sorbate, partial glyceride mixtures of saturatedvegetable fatty acids, water, salts or electrolytes, such as protaminesulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,sodium chloride, zinc salts, colloidal silica, magnesium trisilicate,polyvinyl pyrrolidone, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, wool fat, sugars such aslactose, glucose and sucrose; starches such as corn starch and potatostarch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate; powdered tragacanth;malt; gelatin; talc; excipients such as cocoa butter and suppositorywaxes; oils such as peanut oil, cottonseed oil; safflower oil; sesameoil; olive oil; corn oil and soybean oil; glycols; such a propyleneglycol or polyethylene glycol; esters such as ethyl oleate and ethyllaurate; agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator.

The protein kinase inhibitors or pharmaceutical salts thereof may beformulated into pharmaceutical compositions for administration to asubject as defined herein. These pharmaceutical compositions, whichcomprise an amount of the protein inhibitor effective to treat orprevent a protein kinase-mediated condition and a pharmaceuticallyacceptable carrier, are another embodiment of the present invention.

In one embodiment the present invention is a method of treating orpreventing a protein kinase-mediated disorder in a subject in needthereof, comprising administering to the subject an effective amount ofa compound composition or a pharmaceutically acceptable salt of thepresent invention as described herein. In another embodiment, thepresent invention is the use of an effective amount of a compound,composition or a pharmaceutically acceptable salt described herein fortreating or preventing a disease or disorder, described herein, in asubject in need thereof. In yet another embodiment, the presentinvention is the use of an effective amount of a compound, compositionor a pharmaceutically acceptable salt described herein for themanufacture of a medicament method for the treatment or prevention of adisease or disorder, described herein, in a subject in need thereof. Inone embodiment the protein kinase mediated disease is a protein kinase C(PKC) mediated disease. In another embodiment the protein kinasemediated disease is a protein kinase C theta (PKCtheta)-mediateddisease.

As used herein, the terms “subject”, “patient” and “mammal” are usedinterchangeably. The terms “subject” and “patient” refer to an animal(e.g., a bird such as a chicken, quail or turkey, or a mammal),preferably a mammal including a non-primate (e.g., a cow, pig, horse,sheep, rabbit, guinea pig, rat, cat, dog, and mouse) and a primate(e.g., a monkey, chimpanzee and a human), and more preferably a human.In one embodiment, the subject is a non-human animal such as a farmanimal (e.g., a horse, cow, pig or sheep), or a pet (e.g., a dog, cat,guinea pig or rabbit). In a preferred embodiment, the subject is ahuman.

As used herein, an “effective amount” refers to an amount sufficient toelicit the desired biological response. In the present invention thedesired biological response is to reduce or ameliorate the severity,duration, progression, or onset of a protein kinase-mediated condition,prevent the advancement of a protein kinase-mediated condition, causethe regression of a protein kinase-mediated condition, prevent therecurrence, development, onset or progression of a symptom associatedwith a protein kinase-mediated condition, or enhance or improve theprophylactic or therapeutic effect(s) of another therapy. The preciseamount of compound administered to a subject will depend on the mode ofadministration, the type and severity of the disease or condition and onthe characteristics of the subject, such as general health, age, sex,body weight and tolerance to drugs. It will also depend on the degree,severity and type of protein kinase-mediated condition, and the mode ofadministration. The skilled artisan will be able to determineappropriate dosages depending on these and other factors. Whenco-administered with other agents, e.g., when co-administered with anprotein kinase-mediated condition agent, an “effective amount” of thesecond agent will depend on the type of drug used. Suitable dosages areknown for approved agents and can be adjusted by the skilled artisanaccording to the condition of the subject, the type of condition(s)being treated and the amount of a compound of the invention being used.In cases where no amount is expressly noted, an effective amount shouldbe assumed.

As used herein, the terms “treat”, “treatment” and “treating” refer tothe reduction or amelioration of the progression, severity and/orduration of a protein kinase-mediated condition, or the amelioration ofone or more symptoms (preferably, one or more discernible symptoms) of aprotein kinase-mediated condition resulting from the administration ofone or more therapies (e.g., one or more therapeutic agents such as acompound of the invention). In specific embodiments, the terms “treat”,“treatment” and “treating” refer to the amelioration of at least onemeasurable physical parameter of a protein kinase-mediated condition. Inother embodiments the terms “treat”, “treatment” and “treating” refer tothe inhibition of the progression of a protein kinase-mediatedcondition, either physically by, e.g., stabilization of a discerniblesymptom, physiologically by, e.g., stabilization of a physicalparameter, or both. In other embodiments the terms “treat”, “treatment”and “treating” refer to the reduction or stabilization of a proteinkinase-mediated condition.

As used herein, the terms “prevent”, “prevention” and “preventing” referto the reduction in the risk of acquiring or developing a given proteinkinase-mediated condition, or the reduction or inhibition of therecurrence or a protein kinase-mediated condition. In one embodiment, acompound of the invention is administered as a preventative measure to apatient, preferably a human, having a genetic predisposition to any ofthe conditions, diseases or disorders described herein.

As used herein, the terms, “disease”, “disorder” and “condition” may beused interchangeably here to refer to a protein kinase-mediatedcondition.

In one aspect, the present invention provides a method for treating orlessening the severity of a disease, condition, or disorder where aprotein kinase is implicated in the disease state. In another aspect,the present invention provides a method for treating or lessening theseverity of a kinase disease, condition, or disorder where inhibition ofenzymatic activity is implicated in the treatment of the disease. Inanother aspect, this invention provides a method for treating orlessening the severity of a disease, condition, or disorder withcompounds that inhibit enzymatic activity by binding to the proteinkinase. Another aspect provides a method for treating or lessening theseverity of a kinase disease, condition, or disorder by inhibitingenzymatic activity of the kinase with a protein kinase inhibitor. Insome embodiments, said protein kinase inhibitor is a PKCtheta inhibitor.

The term “protein kinase-mediated condition”, as used herein means anydisease or other deleterious condition in which a protein kinase plays arole. Such conditions include, without limitation, autoimmune diseases,inflammatory diseases, proliferative and hyperproliferative diseases,immunologically-mediated diseases, immuno-deficiency disorders,immunomodulatory or immunosuppressive disorder, bone diseases, metabolicdiseases, neurological and neurodegenerative diseases, cardiovasculardiseases, hormone related diseases, diabetes, allergies, asthma, andAlzheimer's disease.

The term “PKC-mediated condition”, as used herein means any disease orother deleterious condition in which PKC plays a role. Such conditionsinclude, without limitation, those listed above, and in particular,T-cell mediated diseases, including without limitation autoimmunediseases, chronic or acute inflammatory diseases, and proliferative andhyperproliferative diseases.

The term “PKCtheta-mediated condition”, as used herein means any diseaseor other deleterious condition in which PKCtheta plays a role. Suchconditions include, diseases, without limitation, those listed above,and in particular, autoimmune diseases, chronic or acute inflammatorydiseases, and proliferative and hyperproliferative diseases.

As used herein, the term “inflammatory disease” or “inflammatorydisorder” refers to pathological states resulting in inflammation,typically caused by leukocyte infiltration. Examples of such disordersinclude inflammatory skin diseases, including, without limitation,psoriasis and atopic dermatitis; systemic scleroderma and sclerosis;responses associated with inflammatory bowel disease (IBD) (such asCrohn's disease and ulcerative colitis); ischemic reperfusion disordersincluding surgical tissue reperfusion injury, myocardial ischemicconditions such as myocardial infarction, cardiac arrest, reperfusionafter cardiac surgery and constriction after percutaneous transluminalcoronary angioplasty, stroke, and abdominal aortic aneurysms; cerebraledema secondary to stroke; cranial trauma, hypovolemic shock; asphyxia;adult respiratory distress syndrome; acute-lung injury; Behcet'sDisease; dermatomyositis; polymyositis; multiple sclerosis (MS);dermatitis; meningitis; encephalitis; uveitis; osteoarthritis; lupusnephritis; autoimmune diseases such as rheumatoid arthritis (RA),Sjorgen's syndrome, vasculitis; diseases involving leukocyte diapedesis;central nervous system (CNS) inflammatory disorder, multiple organinjury syndrome secondary to septicaemia or trauma; alcoholic hepatitis;bacterial pneumonia; antigen-antibody complex mediated diseasesincluding glomerulonephritis; sepsis; sarcoidosis; immunopathologicresponses to tissue or organ transplantation; inflammations of the lung,including pleurisy, alveolitis, vasculitis, pneumonia, chronicbronchitis, bronchiectasis, diffuse panbronchiolitis, hypersensitivitypneumonitis, idiopathic pulmonary fibrosis (IPF), and cystic fibrosis;etc.

Proliferative or hyperproliferative diseases are characterized byexcessive or abnormal cell proliferation. Such diseases include, withoutlimitation, cancer and myeloproliferative disorders.

The term “cancers” includes, but is not limited to, the followingcancers: epidermoid Oral: Cardiac: Lung: Gastrointestinal: Genitourinarytract: Liver: Bone: Nervous system: Gynecological: Hematologic: Thyroidgland: and Adrenal glands. Hematologic cancers include: blood (myeloidleukemia [acute and chronic], acute lymphoblastic leukemia, chroniclymphocytic leukemia, myeloproliferative diseases, multiple myeloma,myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma[malignant lymphoma] hairy cell; lymphoid disorders; Skin: malignantmelanoma, basal cell carcinoma, squamous cell carcinoma, Karposi'ssarcoma, keratoacanthoma, moles dysplastic nevi, lipoma, angioma,dermatofibroma, keloids, and psoriasis. Thus, the term “cancerous cell”as provided herein, includes a cell afflicted by any one of theabove-identified conditions.

The term “myeloproliferative disorders”, includes disorders such aspolycythemia vera, thrombocythemia, myeloid metaplasia withmyelofibrosis, hypereosinophilic syndrome, juvenile myelomonocyticleukaemia, systemic mast cell disease, and hematopoietic disorders, inparticular, acute-myelogenous leukemia (AML), chronic-myelogenousleukemia (CML), acute-promyelocytic leukemia (APL), and acutelymphocytic leukemia (ALL).

Examples of neurodegenerative diseases include, without limitation,Alzheimer's disease Huntington's disease, Parkinson's disease,AIDS-associated dementia, and bipolar disorder.

In one embodiment the PKCtheta mediated disease includes, withoutlimitation, chronic inflammation, autoimmune diabetes, rheumatoidarthritis (RA), rheumatoid spondylitis, gouty arthritis and otherarthritic conditions, multiple sclerosis (MS), asthma, systemic lupuserythrematosis, adult respiratory distress syndrome, Behcet's disease,psoriasis, chronic pulmonary inflammatory disease, graft versus hostreaction, Crohn's Disease, ulcerative colitis, inflammatory boweldisease (IBD), which includes celiac disease and irritable bowelsyndrome; Alzheimer's disease, T-cell leukaemia, lymphoma, transplantrejection, cancer and pyresis, along with any disease or disorder thatrelates to inflammation and related disorders.

In one embodiment the PKCtheta mediated disease includes—such as,arthritis, rheumatoid arthritis, osteoarthritis, joint inflammation,lupus, multiple sclerosis, asthma, psoriasis, cancer, T-cell lymphomas,leukaemia, diabetes type I or II, and inflammatory bowel diseases,transplant rejection, Crohn's disease and colitis.

Examples of autoimmune diseases include, without limitation, multiplesclerosis, rheumatoid arthritis and irritable bowel disease.

The pharmaceutically acceptable compositions of this invention can beadministered to humans and other animals orally, rectally, parenterally,intracisternally, intravaginally, intraperitoneally, topically (as bypowders, ointments, or drops), bucally, as an oral or nasal spray, orthe like, depending on the severity of the infection being treated.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a compound of the present invention,it is often desirable to slow the absorption of the compound fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the compound thendepends upon its rate of dissolution that, in turn, may depend uponcrystal size and crystalline form. Alternatively, delayed absorption ofa parenterally administered compound form is accomplished by dissolvingor suspending the compound in an oil vehicle. Injectable depot forms aremade by forming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polethylene glycols and the like.

The active compounds can also be in microencapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, eardrops, and eye drops are also contemplated asbeing within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

The compositions of the present invention may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes, but is not limited to, subcutaneous,intravenous, intramuscular, intra-articular, intra-synovial,intrasternal, intrathecal, intrahepatic, intralesional and intracranialinjection or infusion techniques. Preferably, the compositions areadministered orally, intraperitoneally or intravenously.

Sterile injectable forms of the compositions of this invention may beaqueous or oleaginous suspension. These suspensions may be formulatedaccording to techniques known in the art using suitable dispersing orwetting agents and suspending agents. The sterile injectable preparationmay also be a sterile injectable solution or suspension in a non-toxicparenterally-acceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose, any bland fixed oilmay be employed including synthetic mono- or di-glycerides. Fatty acids,such as oleic acid and its glyceride derivatives are useful in thepreparation of injectables, as are natural pharmaceutically-acceptableoils, such as olive oil or castor oil, especially in theirpolyoxyethylated versions. These oil solutions or suspensions may alsocontain a long-chain alcohol diluent or dispersant, such ascarboxymethyl cellulose or similar dispersing agents which are commonlyused in the formulation of pharmaceutically acceptable dosage formsincluding emulsions and suspensions. Other commonly used surfactants,such as Tweens, Spans and other emulsifying agents or bioavailabilityenhancers which are commonly used in the manufacture of pharmaceuticallyacceptable solid, liquid, or other dosage forms may also be used for thepurposes of formulation.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers commonly used include, but arenot limited to, lactose and corn starch. Lubricating agents, such asmagnesium stearate, are also typically added. For oral administration ina capsule form, useful diluents include lactose and dried cornstarch.When aqueous suspensions are required for oral use, the activeingredient is combined with emulsifying and suspending agents. Ifdesired, certain sweetening, flavoring or coloring agents may also beadded.

Alternatively, the pharmaceutical compositions of this invention may beadministered in the form of suppositories for rectal administration.These can be prepared by mixing the agent with a suitable non-irritatingexcipient which is solid at room temperature but liquid at rectaltemperature and therefore will melt in the rectum to release the drug.Such materials include, but are not limited to, cocoa butter, beeswaxand polyethylene glycols.

The pharmaceutical compositions of this invention may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, the pharmaceutical compositions may beformulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this invention include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, the pharmaceutical compositions can be formulatedin a suitable lotion or cream containing the active components suspendedor dissolved in one or more pharmaceutically acceptable carriers.Suitable carriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical compositions may be formulated asmicronized suspensions in isotonic, pH adjusted sterile saline, or,preferably, as solutions in isotonic, pH adjusted sterile saline, eitherwith or without a preservative such as benzylalkonium chloride.Alternatively, for ophthalmic uses, the pharmaceutical compositions maybe formulated in an ointment such as petrolatum.

The pharmaceutical compositions of this invention may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

The dosage regimen utilizing the compounds of Structural Formula I andII can be selected in accordance with a variety of factors including thedisorder being treated and the severity of the disorder; the activity ofthe specific compound employed; the specific composition employed; theage, body weight, general health, sex and diet of the patient; the timeof administration, route of administration, and rate of excretion of thespecific compound employed; the renal and hepatic function of thesubject; and the particular compound or salt thereof employed, theduration of the treatment; drugs used in combination or coincidentalwith the specific compound employed, and like factors well known in themedical arts. The skilled artisan can readily determine and prescribethe effective amount of the compound of Structural Formula I and IIrequired to treat, for example, to prevent, inhibit (fully or partially)or arrest the progress of the disease.

Dosages of the compounds of Structural Formula I and II can range frombetween about 0.01 to about 100 mg/kg body weight/day, about 0.01 toabout 50 mg/kg body weight/day, about 0.1 to about 50 mg/kg bodyweight/day, or about 1 to about 25 mg/kg body weight/day. It isunderstood that the total amount per day can be administered in a singledose or can be administered in multiple dosings such as twice, three orfour times per day.

The compounds for use in the method of the invention can be formulatedin unit dosage form. The term “unit dosage form” refers to physicallydiscrete units suitable as unitary dosage for subjects undergoingtreatment, with each unit containing a predetermined quantity of activematerial calculated to produce the desired therapeutic effect,optionally in association with a suitable pharmaceutical carrier. Theunit dosage form can be for a single daily dose or one of multiple dailydoses (e.g., about 1 to 4 or more times per day). When multiple dailydoses are used, the unit dosage form can be the same or different foreach dose.

An effective amount can be achieved in the method or pharmaceuticalcomposition of the invention employing a compound of Structural FormulaI and II or a pharmaceutically acceptable salt or solvate (e.g.,hydrate) thereof alone or in combination with an additional suitabletherapeutic agent, for example, a cancer-therapeutic agent. Whencombination therapy is employed, an effective amount can be achievedusing a first amount of a compound of Structural Formula I and II or apharmaceutically acceptable salt or solvate (e.g., hydrate) thereof anda second amount of an additional suitable therapeutic agent.

In one embodiment, the compound of Structural Formula I and II and theadditional therapeutic agent, are each administered in an effectiveamount (i.e., each in an amount which would be therapeutically effectiveif administered alone). In another embodiment, the compound ofStructural Formula I and II and the additional therapeutic agent, areeach administered in an amount which alone does not provide atherapeutic effect (a sub-therapeutic dose). In yet another embodiment,the compound of Structural Formula I and II be administered in aneffective amount, while the additional therapeutic agent is administeredin a sub-therapeutic dose. In still another embodiment, the compound ofStructural Formula I and II can be administered in a sub-therapeuticdose, while the additional therapeutic agent, for example, a suitablecancer-therapeutic agent is administered in an effective amount.

As used herein, the terms “in combination” or “coadministration” can beused interchangeably to refer to the use of more than one therapies(e.g., one or more prophylactic and/or therapeutic agents). The use ofthe terms does not restrict the order in which therapies (e.g.,prophylactic and/or therapeutic agents) are administered to a subject.

Coadministration encompasses administration of the first and secondamounts of the compounds of the coadministration in an essentiallysimultaneous manner, such as in a single pharmaceutical composition, forexample, capsule or tablet having a fixed ratio of first and secondamounts, or in multiple, separate capsules or tablets for each. Inaddition, such coadministration also encompasses use of each compound ina sequential manner in either order.

When coadministration involves the separate administration of the firstamount of a compound of Structural Formula I and II and a second amountof an additional therapeutic agent, the compounds are administeredsufficiently close in time to have the desired therapeutic effect. Forexample, the period of time between each administration which can resultin the desired therapeutic effect, can range from minutes to hours andcan be determined taking into account the properties of each compoundsuch as potency, solubility, bioavailability, plasma half-life andkinetic profile. For example, a compound of Structural Formula I and IIand the second therapeutic agent can be administered in any order withinabout 24 hours of each other, within about 16 hours of each other,within about 8 hours of each other, within about 4 hours of each other,within about 1 hour of each other or within about 30 minutes of eachother.

More, specifically, a first therapy (e.g., a prophylactic or therapeuticagent such as a compound of the invention) can be administered prior to(e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeksbefore), concomitantly with, or subsequent to (e.g., 5 minutes, 15minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks,4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) theadministration of a second therapy (e.g., a prophylactic or therapeuticagent such as an anti-cancer agent) to a subject.

It is understood that the method of coadministration of a first amountof a compound of Structural Formula I and II and a second amount of anadditional therapeutic agent can result in an enhanced or synergistictherapeutic effect, wherein the combined effect is greater than theadditive effect that would result from separate administration of thefirst amount of the compound of Structural Formula I and II and thesecond amount of the additional therapeutic agent.

As used herein, the term “synergistic” refers to a combination of acompound of the invention and another therapy (e.g., a prophylactic ortherapeutic agent), which is more effective than the additive effects ofthe therapies. A synergistic effect of a combination of therapies (e.g.,a combination of prophylactic or therapeutic agents) permits the use oflower dosages of one or more of the therapies and/or less frequentadministration of said therapies to a subject. The ability to utilizelower dosages of a therapy (e.g., a prophylactic or therapeutic agent)and/or to administer said therapy less frequently reduces the toxicityassociated with the administration of said therapy to a subject withoutreducing the efficacy of said therapy in the prevention, management ortreatment of a disorder. In addition, a synergistic effect can result inimproved efficacy of agents in the prevention, management or treatmentof a disorder. Finally, a synergistic effect of a combination oftherapies (e.g., a combination of prophylactic or therapeutic agents)may avoid or reduce adverse or unwanted side effects associated with theuse of either therapy alone.

The presence of a synergistic effect can be determined using suitablemethods for assessing drug interaction. Suitable methods include, forexample, the Sigmoid-Emax equation (Holford, N. H. G. and Scheiner, L.B., Clin. Pharmacokinet. 6: 429-453 (1981)), the equation of Loeweadditivity (Loewe, S. and Muischnek, H., Arch. Exp. Pathol Pharmacol.114: 313-326 (1926)) and the median-effect equation (Chou, T. C. andTalalay, P., Adv. Enzyme Regul. 22: 27-55 (1984)). Each equationreferred to above can be applied with experimental data to generate acorresponding graph to aid in assessing the effects of the drugcombination. The corresponding graphs associated with the equationsreferred to above are the concentration-effect curve, isobologram curveand combination index curve, respectively.

In some embodiments, said additional therapeutic agent is selected froma cancer-therapeutic agent, such as, an anti-cancer agent, ananti-proliferative agent, or a chemotherapeutic agent.

In some embodiments, said additional therapeutic agent is selected fromcamptothecin, the MEK inhibitor: U0126, a KSP (kinesin spindle protein)inhibitor, adriamycin, interferons, and platinum derivatives, such asCisplatin.

In other embodiments, said additional therapeutic agent is selected fromtaxanes; inhibitors of bcr-abl (such as Gleevec, dasatinib, andnilotinib); inhibitors of EGFR (such as Tarceva and Iressa); DNAdamaging agents (such as cisplatin, oxaliplatin, carboplatin,topoisomerase inhibitors, and anthracyclines); and antimetabolites (suchas AraC and 5-FU).

In yet other embodiments, said additional therapeutic agent is selectedfrom camptothecin, doxorubicin, idarubicin, Cisplatin, taxol, taxotere,vincristine, tarceva, the MEK inhibitor, U0126, a KSP inhibitor,vorinostat, Gleevec, dasatinib, and nilotinib.

In another embodiment, said additional therapeutic agent is selectedfrom Her-2 inhibitors (such as Herceptin); HDAC inhibitors (such asvorinostat), VEGFR inhibitors (such as Avastin), c-KIT and FLT-3inhibitors (such as sunitinib), BRAF inhibitors (such as Bayer's BAY43-9006) MEK inhibitors (such as Pfizer's PD0325901); and spindlepoisons (such as Epothilones and paclitaxel protein-bound particles(such as Abraxane®).

Other therapies or anticancer agents that may be used in combinationwith the inventive agents of the present invention include surgery,radiotherapy (in but a few examples, gamma-radiation, neutron beamradiotherapy, electron beam radiotherapy, proton therapy, brachytherapy,and systemic radioactive isotopes, to name a few), endocrine therapy,biologic response modifiers (interferons, interleukins, and tumornecrosis factor (TNF) to name a few), hyperthermia and cryotherapy,agents to attenuate any adverse effects (e.g., antiemetics), and otherapproved chemotherapeutic drugs, including, but not limited to,alkylating drugs (mechlorethamine, chlorambucil, Cyclophosphamide,Melphalan, Ifosfamide), antimetabolites (Methotrexate), purineantagonists and pyrimidine antagonists (6-Mercaptopurine,5-Fluorouracil, Cytarabile, Gemcitabine), spindle poisons (Vinblastine,Vincristine, Vinorelbine, Paclitaxel), podophyllotoxins (Etoposide,Irinotecan, Topotecan), antibiotics (Doxorubicin, Bleomycin, Mitomycin),nitrosoureas (Carmustine, Lomustine), inorganic ions (Cisplatin,Carboplatin), enzymes (Asparaginase), and hormones (Tamoxifen,Leuprolide, Flutamide, and Megestrol), Gleevec™, adriamycin,dexamethasone, and cyclophosphamide.

A compound of the instant invention may also be useful for treatingcancer in combination with any of the following therapeutic agents:abarelix (Plenaxis Depot®); aldesleukin (Prokine®); Aldesleukin(Proleukin®); Alemtuzumabb (Campath®); alitretinoin (Panretin®);allopurinol (Zyloprim®); altretamine (Hexalen®); amifostine (Ethyol®);anastrozole (Arimidex®); arsenic trioxide (Trisenox®); asparaginase(Elspar®); azacitidine (Vidaza®); bevacuzimab (Avastin®); bexarotenecapsules (Targretin®); bexarotene gel (Targretin®); bleomycin(Blenoxane®); bortezomib (Velcade®); busulfan intravenous (Busulfex®);busulfan oral (Myleran®); calusterone (Methosarb®); capecitabine(Xeloda®); carboplatin (Paraplatin®); carmustine (BCNU®, BiCNU®);carmustine (Gliadel®); carmustine with Polifeprosan 20 Implant (GliadelWafer®); celecoxib (Celebrex®); cetuximab (Erbitux®); chlorambucil(Leukeran®); cisplatin (Platinol®); cladribine (Leustatin®, 2-CdA®);clofarabine (Clolar®); cyclophosphamide (Cytoxan®, Neosar®);cyclophosphamide (Cytoxan Injection®); cyclophosphamide (CytoxanTablet®); cytarabine (Cytosar-U®); cytarabine liposomal (DepoCyt®);dacarbazine (DTIC-Dome®); dactinomycin, actinomycin D (Cosmegen®);Darbepoetin alfa (Aranesp®); daunorubicin liposomal (DanuoXome®);daunorubicin, daunomycin (Daunorubicin®); daunorubicin, daunomycin(Cerubidine®); Denileukin diftitox (Ontak®); dexrazoxane (Zinecard®);docetaxel (Taxotere®); doxorubicin (Adriamycin PFS®); doxorubicin(Adriamycin®, Rubex®); doxorubicin (Adriamycin PFS Injection®);doxorubicin liposomal (Doxil®); dromostanolone propionate(Dromostanolone®); dromostanolone propionate (Masterone Injection®);Elliott's B Solution (Elliott's B Solution®); epirubicin (Ellence®);Epoetin alfa (Epogen®); erlotinib (Tarceva®); estramustine (Emcyt®);etoposide phosphate (Etopophos®); etoposide, VP-16 (Vepesid®);exemestane (Aromasin®); Filgrastim (Neupogen®); floxuridine(intraarterial) (FUDR®); fludarabine (Fludara®); fluorouracil, 5-FU(Adrucil®); fulvestrant (Faslodex®); gefitinib (Iressa®); gemcitabine(Gemzar®); gemtuzumab ozogamicin (Mylotarg®); goserelin acetate (ZoladexImplant®); goserelin acetate (Zoladex®); histrelin acetate (HistrelinImplant®); hydroxyurea (Hydrea®); Ibritumomab Tiuxetan (Zevalin®);idarubicin (Idamycin®); ifosfamide (IFEX®); imatinib mesylate(Gleevec®); interferon alfa 2a (Roferon A®); Interferon alfa-2b (IntronA®); irinotecan (Camptosar®); lenalidomide (Revlimid®); letrozole(Femara®); leucovorin (Wellcovorin®, Leucovorin®); Leuprolide Acetate(Eligard®); levamisole (Ergamisol®); lomustine, CCNU (CeeBU®);meclorethamine, nitrogen mustard (Mustargen®); megestrol acetate(Megace®); melphalan, L-PAM (Alkeran®); mercaptopurine, 6-MP(Purinethol®); mesna (Mesnex®); mesna (Mesnex Tabs®); methotrexate(Methotrexate®); methoxsalen (Uvadex®); mitomycin C (Mutamycin®);mitotane (Lysodren®); mitoxantrone (Novantrone®); nandrolonephenpropionate (Durabolin-50®); nelarabine (Arranon®); Nofetumomab(Verluma®); Oprelvekin (Neumega®); oxaliplatin (Eloxatin®); paclitaxel(Paxene®); paclitaxel (Taxol®); paclitaxel protein-bound particles(Abraxane®); palifermin (Kepivance®); pamidronate (Aredia®); pegademase(Adagen (Pegademase Bovine)®); pegaspargase (Oncaspar®); Pegfilgrastim(Neulasta®); pemetrexed disodium (Alimta®); pentostatin (Nipent®);pipobroman (Vercyte®); plicamycin, mithramycin (Mithracin®); porfimersodium (Photofrin®); procarbazine (Matulane®); quinacrine (Atabrine®);Rasburicase (Elitek®); Rituximab (Rituxan®); sargramostim (Leukine®);Sargramostim (Prokine®); sorafenib (Nexavar®); streptozocin (Zanosar®);sunitinib maleate (Sutent®); talc (Sclerosol®); tamoxifen (Nolvadex®);temozolomide (Temodar®); teniposide, VM-26 (Vumon®); testolactone(Teslac®); thioguanine, 6-TG (Thioguanine®); thiotepa (Thioplex®);topotecan (Hycamtin®); toremifene (Fareston®); Tositumomab (Bexxar®);Tositumomab/I-131 tositumomab (Bexxar®); Trastuzumab (Herceptin®);tretinoin, ATRA (Vesanoid®); Uracil Mustard (Uracil Mustard Capsules®);valrubicin (Valstar®); vinblastine (Velban®); vincristine (Oncovin®);vinorelbine (Navelbine®); zoledronate (Zometa®) and vorinostat(Zolinza®).

For a comprehensive discussion of updated cancer therapies see,http://www.nci.nih.gov/, a list of the FDA approved oncology drugs athttp://www.fda.gov/cder/cancer/druglistframe.htm, and The Merck Manual,Seventeenth Ed. 1999, the entire contents of which are herebyincorporated by reference.

Other examples of agents the compounds of this invention may also becombined with include, without limitation: treatments for Alzheimer'sDisease such as Aricept® and Excelon®; treatments for Parkinson'sDisease such as L-DOPA/carbidopa, entacapone, ropinrole, pramipexole,bromocriptine, pergolide, trihexephendyl, and amantadine; agents fortreating Multiple Sclerosis (MS) such as beta interferon (e.g., Avonex®and Rebif®), Copaxone®, and mitoxantrone; treatments for asthma such asalbuterol and Singulair®; agents for treating schizophrenia such aszyprexa, risperdal, seroquel, and haloperidol; anti-inflammatory agentssuch as corticosteroids, TNF blockers, IL-1 RA, azathioprine,cyclophosphamide, and sulfasalazine; immunomodulatory andimmunosuppressive agents such as cyclosporin, tacrolimus, rapamycin,mycophenolate mofetil, interferons, corticosteroids, cyclophophamide,azathioprine, and sulfasalazine; neurotrophic factors such asacetylcholinesterase inhibitors, MAO inhibitors, interferons,anti-convulsants, ion channel blockers, riluzole, and anti-Parkinsonianagents; agents for treating cardiovascular disease such asbeta-blockers, ACE inhibitors, diuretics, nitrates, calcium channelblockers, and statins; agents for treating liver disease such ascorticosteroids, cholestyramine, interferons, and anti-viral agents;agents for treating blood disorders such as corticosteroids,anti-leukemic agents, and growth factors; and agents for treatingimmunodeficiency disorders such as gamma globulin.

As inhibitors of protein kinases, the compounds and compositions of thisinvention are also useful in biological samples. One aspect of theinvention relates to inhibiting protein kinase activity in a biologicalsample, which method comprises contacting said biological sample with acompound of formula I and II or a composition comprising said compound.The term “biological sample”, as used herein, means an in vitro or an exvivo sample, including, without limitation, cell cultures or extractsthereof; biopsied material obtained from a mammal or extracts thereof;and blood, saliva, urine, feces, semen, tears, or other body fluids orextracts thereof.

Inhibition of protein kinase activity in a biological sample is usefulfor a variety of purposes that are known to one of skill in the art.Examples of such purposes include, but are not limited to, bloodtransfusion, organ-transplantation, and biological specimen storage.

Another aspect of this invention relates to the study of protein kinasesin biological and pathological phenomena; the study of intracellularsignal transduction pathways mediated by such protein kinases; and thecomparative evaluation of new protein kinase inhibitors. Examples ofsuch uses include, but are not limited to, biological assays such asenzyme assays and cell-based assays.

The activity of the compounds as protein kinase inhibitors may beassayed in vitro, in vivo or in a cell line. In vitro assays includeassays that determine inhibition of either the kinase activity or ATPaseactivity of the activated kinase. Alternate in vitro assays quantitatethe ability of the inhibitor to bind to the protein kinase and may bemeasured either by radiolabelling the inhibitor prior to binding,isolating the inhibitor/kinase complex and determining the amount ofradiolabel bound, or by running a competition experiment where newinhibitors are incubated with the kinase bound to known radioligands.Detailed conditions for assaying a compound utilized in this inventionis set forth in the Examples below.

Another aspect of this invention relates to the use of the compoundsdescribed here (in particular those with moderate observed affinity forbiochemical targets (IC50 1-10 μM)) as start points for chemistryoptimization. In particular, one aspect of this invention relates toroutine inhibition studies against a target enzyme for chemicaloptimization.

Another aspect of this invention relates to the use of the compoundsdescribed herein for crystallography (in particular those with moderateobserved affinity for biochemical targets): In particular, the oneaspect of this invention relates to the generation of co-complex crystalstructures with compounds described herein.

Another aspect of this invention relates to the use of the compoundsdescribed herein as chemical tools to probe target biology in vitro andin vivo: In particular inhibitors with moderate affinity in biochemicalassays can be used to probe the biological impact of inhibiting a targetenzyme in cells and in whole animal models of disease.

Another aspect of the invention provides a method for modulating enzymeactivity by contacting a compound of formula I, and II with a proteinkinase.

Abbreviations

The following abbreviations are used:

DMSO dimethyl sulfoxide

TCA trichloroacetic acid

ATP adenosine triphosphate

BSA bovine serum albumin

DTT dithiothreitol

MOPS 4-morpholinepropanesulfonic acid

NMR nuclear magnetic resonance

HPLC high performance liquid chromatography

LCMS liquid chromatography-mass spectrometry

TLC thin layer chromatography

Rt retention time

In some embodiments, the compounds of this invention are represented inTable 1. In certain embodiments, the variables used herein are asdefined in the specific embodiments as shown in Table 1.

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45or a pharmaceutically acceptable salt thereofGeneral Synthetic Methodology

The compounds of this invention may be prepared in light of thespecification using steps generally known to those of ordinary skill inthe art. Those compounds may be analyzed by known methods, including butnot limited to LCMS (liquid chromatography mass spectrometry) HPLC andNMR (nuclear magnetic resonance). It should be understood that thespecific conditions shown below are only examples, and are not meant tolimit the scope of the conditions that can be used for making compoundsof this invention. Instead, this invention also includes conditions thatwould be apparent to those skilled in that art in light of thisspecification for making the compounds of this invention. Unlessotherwise indicated, all variables in the following schemes are asdefined herein. General Schemes:

EXAMPLES

Mass spec. samples were analyzed on a MicroMass Quattro Micro massspectrometer operated in single MS mode with electrospray ionization.Samples were introduced into the mass spectrometer using chromatography.Mobile phase for all mass spec. analyses consisted of 10 mM pH 7ammonium acetate and a 1:1 acetonitrile-methanol mixture. Method A:Column gradient conditions were 5%-100% acetonitrile-methanol over 3.5mins gradient time and 4.8 mins run time on an ACE5C8 3.0×75 mm column.Flow rate was 1.2 ml/min. Method B: Column gradient were 5%-100%acetonitrile-methanol over 10 mins gradient time and 12 mins run time ona ACE5C8 4.6×150 mm column. Flow rate was 1.5 mL/min. As used herein,the term “Rt(min)” refers to the LCMS retention time, in minutes,associated with the compound. Unless otherwise indicated, the LCMSmethod utilized to obtain the reported retention time is as detailedabove. If the Rt(min) is <5 min method A was used, if the Rt(min) is >5min then method B was used.

1H-NMR spectra were recorded at 400 MHz using a Bruker DPX 400instrument.

The following compounds of formula I, and II can be prepared andanalyzed as follows:

Scheme I

Scheme I above shows a general route for the preparation of compounds offormula E, wherein the variables, are as defined herein and N(W)₂ formsthe piperazine/pyrrolodine ring as defined herein. The weinreb amide Ais coupled with compound B in the presence of n-butyl lithium orGrignard reagent to form a compound of formula C. Compound C is thenheated in the presence of hydrazine to yield intermediate D. Thecompound of formula D is displaced by an optionally protected amine inthe presence of suitable base, such as, potassium carbonate,diisopropylethylamine (DIPEA), triethylamine,1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) etc., in a suitable solvent,such as for example, dimethylformamide, dimethylsulfoxide (DMSO),n-butanol (n-Bu-OH) etc., at about 70° C. to about 110° C., about 80° C.to about 100° C., about 90° C. to about 100° C. to form an aminesubstituted heteroaroyl pyrazolopyridine. Alternatively the displacementcan be perform using Buchwall type condition using Pd as catalyst and aseries of bases and ligands well known by those skilled in the art

Scheme II above shows a general route for the preparation of compoundsof formula K, wherein the variables, are as defined herein. The Weinrebamide A is coupled with compound G in the presence of lithiumdiisopropylamide (LDA) to form a compound of formula H. Compound H isthen treated with hydrazine to yield intermediate I. The compound offormula I is displaced by an optionally protected amine in the presenceof suitable base, such as, potassium carbonate, diisopropylethylamine(DIPEA), triethylamine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) etc.,in a suitable solvent, such as for example, dimethylformamide,dimethylsulfoxide (DMSO), n-butanol (n-Bu-OH) etc., at about 70° C. toabout 110° C. to form an amine substituted heteroaroyl pyrazolopyridine.Alternatively the displacement can be performed using Buchwald typecondition using Pd as catalyst and a series of bases and ligands wellknown by those skilled in the art.

Scheme III above shows a general route for the preparation of compoundsof formula O, wherein the variables, are as defined herein. The boronatederivative L is coupled with pyridine derivative L in the presence of Pdas catalyst in a Suzuki coupling reaction to form a compound of formulaN. Compound N is then displaced by an optionally protected amine in thepresence of suitable base, such as, potassium carbonate,diisopropylethylamine (DIPEA), triethylamine,1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) etc., in a suitable solvent,such as for example, dimethylformamide, dimethylsulfoxide (DMSO),n-butanol (n-Bu-OH), N-methylpyrrolidinone (NMP) etc., at about 70° C.to about 110° C. to form an amine substituted heteroaroylpyrazolopyridine. Alternatively the displacement can be performed usingBuchwald type condition using Pd as catalyst and a series of bases andligands well known by those skilled in the art. Final deprotection yieldcompounds of general formula 0.

Scheme IV above shows a general route for the preparation of compoundsof formula R, wherein the variables, are as defined herein. M isdisplaced with an optionally protected amine in the presence of asuitable as such as potassium carbonate, diisopropylethylamine (DIPEA),triethylamine, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) etc., in asuitable solvent, such as for example, dimethylformamide,dimethylsulfoxide (DMSO), n-butanol (n-Bu-OH), N-methylpyrrolidinone(NMP) at about 100° C. to about 130° C. to form an amine substitutedpyridine P. The boronate derivative L is coupled with pyridinederivative P in the presence of Pd as catalyst in a Suzuki couplingreaction to form a compound of formula Q. Final deprotection yieldcompounds of general formula R.

Example 1 Compound 1 (R)-(+)-3-methyl-2-(pyrazin-2-yl)butan-2-ol

A solution of 2,2,6,6-tetramethylpiperidine (282.2 g, 337.2 mL, 1.998mol) in dry THF (1.400 L) was cooled to −35° C. under nitrogen. nBuLi(2.5M in hexanes) (799.2 mL of 2.5 M, 1.998 mol) was added over 30minutes, keeping the internal temperature between −25 and −35° C. Themixture was allowed to warm to 0° (+/− 1°) and stirred for 10 minutes.The mixture was then cooled to −78° C. and held at this temperature for20 minutes. A solution of pyrazine (80 g, 998.9 mmol) in dry THF (200.0mL) was then cannulated into the lithium amide mixture, keeping theinternal temperature below −70° C. The deep red mixture was stirred for20 minutes, then 3-methylbutan-2-one (430.1 g, 534.3 mL, 4.994 mol)(cooled in a −78° C. bath) was added as rapidly as possible, keeping theinternal temperature below −60°. The mixture was then stirred at −78° C.(+/−5° C.) for 2 hrs. Lc/Ms after this time showed no pyrazineremaining. The reaction was quenched at −78° C. by slow addition of 2MHCl (2 L). Ethyl acetate (800 mL) was then added. The mixture wasallowed to warm to ambient and the organic phase separated. The mixturewas very dark but the phases were easily separated. The aqueous phasewas extracted with ethyl acetate (2×500 ml). The combined organics werewashed with sat NaHCO3 (800 mL), then water (1 L) then brine (500 ml).The mixture was then concentrated. The residue was dissolved in petrol(1.2 L). Some solid was present. MgSO4 was added and the mixture dried,filtered and concentrated. The crude was a dark red oil (260 g). Thecrude was purified on silica gel, eluting with 0-50% ethylacetate/petrol). The title compound was obtained as a pale yellow oil(103.2 g, 62%); ¹H NMR (CDCl₃) 0.74 (3H, d), 1.01 (3H, d), 2.07 (1H, m),4.19 (1H, s), 8.52 (2H, s), 8.73 (1H, s); MS ES(+) 167.35 (M+1).

The (R)-(+)- enatiomer was separated by chiral SFC chromatography usinga cellulose-2 column (Phenomenex) (10% AcCN, 5 ml/min, 100 bar, 35° C.,220 nm; retention time 1.54 min) [ ]_(D)+23.8° (c=5.15, EtOH).

(2R)-3-methyl-2-(piperazin-2-yl)butan-2-ol

Platinum oxide (4.673 g, 20.58 mmol) was placed in a Parr bottle.Methanol (140 mL) was added under an atmosphere of nitrogen followed by(R)-(+)-3-methyl-2-(pyrazin-2-yl)butan-2-ol (17.1 g, 102.9 mmol). Thereaction mixture was shaken in a PARR hydrogenator overnight with ahydrogen pressure of 60 psi. Lc/Ms after this time indicate that thereaction was complete. The suspension was filtered through a pad ofcelite under a nitrogen atmosphere, rinsing with methanol. The crudemixture was concentrated in vacuo. The residue was re-dissolved indichloromethane, dried over MgSO4, filtered and concentrated in vacuo toafford the title compound as viscous light yellow oil (16.6 g, 93%); ¹HNMR (CDCl₃) 0.82-1.08 (9H, m), 1.81 (1H, m), 2.20-3.17 (10H, m); MSES(+) 173.0 (M+1).

4-(tert-butyldimethylsilyl)-2,3,5-trifluoropyridine

A solution of diisopropylamine (98.85 g, 136.9 mL, 976.9 mmol) in THF(1.350 L) was cooled to −65° C. n-BuLi (2.5 M in hexanes) (375.8 mL of2.5 M, 939.4 mmol) was added dropwise via cannula over 1 h at such arate as to maintain reaction temperature below −60° C. Once the additionwas complete the cooling bath was removed and the reaction mixture wasallowed to warm up to 0° C. The reaction mixture was stirred for 15 minat 0° C., then re-cooled to −78° C. 2,3,5-trifluoropyridine (100 g,751.5 mmol) was added dropwise via cannula over 20 min at such a rate tomaintain the reaction temperature below −69° C. The reaction mixture wasstirred for 45 min at −78° C. during which time the solution turnedorange brown. A solution of tert-butyl-chloro-dimethyl-silane (147.2 g,976.9 mmol) in THF (150 mL) was then added via cannula over 30 min. Thereaction mixture was stirred at −78° C. for 90 minutes during which timethe solution darkened. Lc/Ms after this time indicated that the reactionwas complete. A saturated ammonium chloride solution (300 ml) was thenadded and mixture was allowed to warm up to RT. The reaction mixture wasdiluted with water (100 ml) and extracted with EtOAc (1.5 L then 2×500ml). The combined organics were washed with saturated NaHCO (500 ml) andbrine (400 ml). The crude mixture was partially concentrated in vacuo,dried over magnesium sulfate, filtered and concentrated in vacuo to anoil. The crude was purified by flash chromatography (CombiFlashCompanion XL, 1.5 kg column, 0-20% ethyl acetate in petroleum ether).This afforded the title compound as a colourless oil (136.2 g, 73%); ¹HNMR (CDCl₃) 0.34 (6H, s), 0.89 (9H, s), 7.73 (1H, s); MS ES(+) 248.25(M+1).

(4-(tert-butyldimethylsilyl)-3,5,6-trifluoropyridin-2-yl)(2-fluoropyridin-3-yl)methanone

A solution of diisopropylamine (66.78 g, 92.49 mL, 659.9 mmol) in THF(1.360 L) was cooled to −20° C. under nitrogen. nBuLi (2.5M in hexane)(253.0 mL of 2.5 M, 632.4 mmol) was added dropwise via cannula at such arate as to maintain the internal temperature below −15° C. The solutionwas warmed to 0° C. then immediately re-cooled to −90° C.Tert-butyl-dimethyl-(2,3,5-trifluoro-4-pyridyl)silane (136 g, 549.9mmol) was added dropwise via cannula at such a rate as to maintain theinternal temperature below −85° C. After complete addition the solutionbecame an orange suspension. The reaction mixture was stirred at −85° C.for 1 h then 2-fluoro-N-methoxy-N-methylpyridine-3-carboxamide (116.5 g,632.4 mmol) was added dropwise over 1 h keeping the internal temperaturebelow −85° C. The mixture turned dark green then dark red duringaddition. The mixture was stirred at −85° C. for 45 min after which timeLc/Ms indicated the reaction was complete. The cooling bath was removedand the mixture warmed to −50° C. Saturated ammonium chloride solutionNH4Cl (300 mL) was added and the mixture allowed to warm up to RT. Themixture was diluted with EtOAc (2.5 L). The aqueous phase was separatedand extracted with more EtOAc (500 ml). The combined organics werewashed with brine and partially concentrated in vacuo. The solution wasdried over magnesium sulfate, filtered and concentrated in vacuo to abrown oil. The crude was purified on silica gel, eluting with 0-20%ethyl acetate in petroleum ether. This gave the title compound as ayellow oil which solidified on standing (159.6 g, 78%); ¹H NMR (CDCl₃)0.35 (6H, s), 0.88 (9H, s), 7.29 (1H, m), 8.13 (1H, m), 8.37 (1H, m);¹⁹F NMR (decoupled) −112.47, −106.7, −89.3, −61.95; MS ES (+) 371.14(M+1).

3-(4-(tert-butyldimethylsilyl)-3,5,6-trifluoropyridin-2-yl)-1H-pyrazolo[3,4-b]pyridine

[4-[tert-butyl(dimethyl)silyl]-3,5,6-trifluoro-2-pyridyl]-(2-fluoro-3-pyridyl)methanone(159 g, 429.2 mmol) was dissolved in dioxane (1.6 L) and calciumcarbonate (85.91 g, 858.4 mmol) was added. The reaction was in an icebath and hydrazine hydrate (107.4 g, 104.4 mL, 2.146 mol) was addeddropwise over 30 minutes. The resultant mixture was stirred overnight,after which time the thick suspension became a heterogeneous redsolution. The reaction mixture was filtered through a pad of Celite,washing copiously with EtOAc/MeOH 7:1 (2×500 mL) and a small amount ofwater (100 mL). The filtrate was partitioned between EtOAc (1 L) andwater (500 ml). The organic phase was washed with a sat aq NaHCO3 (500mL). The combined aqueous layers were back-extracted with EtOAc (2×200ml) and the combined organics were washed with brine (400 ml), driedover MgSO4 and filtered and concentrated to an orange solid. The solidwas triturated with dichloromethane and filtered, washing withdichloromethane and petrol. This gave the title compound as an off-whitesolid (103.8 g). The filtrate was concentrated under reduced pressure.The resulting solid was again triturated and filtered, washing with morepetrol to give an off-white solid (36.7 g). The title compound wasobtained as an off-white solid (total weight 140.5 g, 90%); ¹H NMR(CDCl₃) 0.55 (6H, s), 1.02 (9H, s), 7.21 (1H, m), 8.69 (1H, m), 8.91(1H, s), 11.69 (1H, brs); MS ES (+) 365.18 (M+1).

3-(4-(tert-butyldimethylsilyl)-3,5-difluoro-6-((S)-3-((R)-3-methyl-2-(trimethylsilyloxy)butan-2-yl)piperazin-1-yl)pyridin-2-yl)-1H-pyrazolo[3,4-b]pyridine

A 500 ml glass pressure vessel was charged withtert-butyl-dimethyl-[2,3,5-trifluoro-6-(1H-pyrazolo[3,4-b]pyridin-3-yl)-4-pyridyl]silane(26.80 g, 73.53 mmol), (2R)-3-methyl-2-piperazin-2-yl-butan-2-ol (19 g,110.3 mmol), THF (53 mL) and imidazol-1-yl-trimethyl-silane (51.57 g,53.72 mL, 367.7 mmol). An identical experiment was run in parallel. Thecontainers was sealed and the mixture was heated and stirred at 95° C.overnight. Lc/Ms after this time showed a mixture consisting of 73% ofthe desired (S,R)-diastereoisomer and 18% of the (R,R)-diastereoisomer.Both reaction mixtures were cooled, combined and diluted with EtOAc (500ml). The solution was washed with saturated sodium bicarbonate (2×200ml). The combined aqueous layers are extracted 3 times with ethylacetate (200 ml). The combined organics were washed with brine (200 ml),dried over MgSO4, filtered and concentrated in vacuo to give 163.874 gof viscous brown oil. The diastereoisomers were separated on silica gel,eluting with 10-95% ethyl acetate in petroleum ether containing 2%triethylamine) This gave the required (S,R)-diastereoisomer as an orangefoam (60.1 g, 69%); ¹H NMR (CDCl₃) 0.001 (9H, s), 0.32 (6H, s),0.76-0.84 (9H, m), 1.11 (4H, m), 1.45 (4H, m), 1.72 (1H, m), 1.90 (1H,s), 2.61 (1H, m), 2.74 (2H, m), 2.97 (1H, m), 3.11 (1H, m), 3.64 (1H,m), 3.82 (1H, m), 3.96 (1H, m), 7.09 (1H, m), 8.48 (1H, m), 8.74 (1H,m), 11.31 (1H, brs); ¹⁹F NMR (decoupled) −113.45, −108.39; MS ES(+)589.3 (M+1). The (R,R)-diastereoisomer was isolated as an orange foam(22%); ¹H NMR (CDCl₃) 0.001 (9H, s), 0.33 (6H, s), 0.72-0.88 (10H, m),1.07 (3H, m), 1.45 (5H, m), 1.86 (1H, m), 2.54 (1H, m), 2.74 (2H, m),2.94 (1H, m), 3.07 (1H, m), 3.63 (1H, m), 3.73 (1H, m), 3.96 (1H, m),7.04 (1H, m), 8.46 (1H, m), 8.69 (1H, m), 11.35 (1H, brs); ¹⁹F NMR(decoupled) −113.78, −108.4; MS ES(+) 589.3 (M+1).

(R)-2-((S)-4-(3,5-difluoro-6-(1H-pyrazolo[3,4-b]pyridin-3-yl)pyridin-2-yl)piperazin-2-yl)-3-methylbutan-2-ol

tert-butyl-[2-[(3S)-3-[(1R)-1,2-dimethyl-1-trimethylsilyloxy-propyl]piperazin-1-yl]-3,5-difluoro-6-(1H-pyrazolo[3,4-b]pyridin-3-yl)-4-pyridyl]-dimethyl-silane(60 g, 101.9 mmol) was dissolved in THF (216.0 mL) at room temperature.TBAF in THF (214.0 mL of 1 M, 214.0 mmol) was added dropwise via cannulato the brown solution over 45 minutes. The reaction was stirredovernight, after which time the deprotection was complete by Lc/Ms. Themixture was diluted with EtOAc (600 ml) and washed with saturatedaqueous NaHCO3 (100 ml then 2×200 ml). The organic phase was furtherwashed with a 2:3 mixture of saturated brine:water (100 ml) until noremaining Bu4N+ salts were observed Lc/Ms analysis. The organic phasewas dried over MgSO4, filtered and concentrated in vacuo to give a brownsolid. The solid was triturated with diethyl ether, filtered and driedto give a orange brown solid (34.5 g, 84%). The solid was recrystallisedfrom isopropanol. This afforded the title compound as a beige solid (23g); ¹H NMR (d6-DMSO) 0.84 (3H, d), 0.90 (3H, d), 1.06 (3H, s), 1.83 (2H,m), 2.74 (2H, d), 2.85 (2H, d), 3.09 (1H, d), 3.78 (1H, d), 4.04 (1H,d), 4.13 (1H, s), 7.30 (1H, dd), 7.92 (1H, dd), 8.60 (1H, dd), 8.77 (1H,dd), 13.99 (1H, brs); ¹⁹F NMR (decoupled) −128.5, −124.1; MS ES(+) 403.0(M+1).

A second crop was obtained from the filtrate (5.3 g) and had identicalspectroscopic data.

Example 2 Compound 9

(R)-(+)-3-methyl-2-(pyrazin-2-yl)butan-2-ol

A solution of 2,2,6,6-tetramethylpiperidine (282.2 g, 337.2 mL, 1.998mol) in dry THF (1.400 L) was cooled to −35° C. under nitrogen. nBuLi(2.5M in hexanes) (799.2 mL of 2.5 M, 1.998 mol) was added over 30minutes, keeping the internal temperature between −25 and −35° C. Themixture was allowed to warm to 0° (+/−1°) and stirred for 10 minutes.The mixture was then cooled to −78° C. and held at this temperature for20 minutes. A solution of pyrazine (80 g, 998.9 mmol) in dry THF (200.0mL) was then cannulated into the lithium amide mixture, keeping theinternal temperature below −70° C. The deep red mixture was stirred for20 minutes, then 3-methylbutan-2-one (430.1 g, 534.3 mL, 4.994 mol)(cooled in a −78° C. bath) was added as rapidly as possible, keeping theinternal temperature below −60°. The mixture was then stirred at −78° C.(+/−5° C.) for 2 hrs. Lc/Ms after this time showed no pyrazineremaining. The reaction was quenched at −78° C. by slow addition of 2MHCl (2 L). Ethyl acetate (800 mL) was then added. The mixture wasallowed to warm to ambient and the organic phase separated. The mixturewas very dark but the phases were easily separated. The aqueous phasewas extracted with ethyl acetate (2×500 ml). The combined organics werewashed with sat NaHCO3 (800 ml), then water (1 L) then brine (500 ml).The mixture was then concentrated. The residue was dissolved in petrol(1.2 L). Some solid was present. MgSO4 was added and the mixture dried,filtered and concentrated. The crude was a dark red oil (260 g). Thecrude was purified on silica gel, eluting with 0-50% ethylacetate/petrol). The title compound was obtained as a pale yellow oil(103.2 g, 62%); ¹H NMR (CDCl₃) 0.74 (3H, d), 1.01 (3H, d), 2.07 (1H, m),4.19 (1H, s), 8.52 (2H, s), 8.73 (1H, s); MS ES(+) 167.35 (M+1).

The (R)-(+)-enatiomer was separated by chiral SFC chromatography using acellulose-2 column (Phenomenex) (10% AcCN, 5 ml/min, 100 bar, 35° C.,220 nm; retention time 1.54 min) [ ]_(D)+23.8° (c=5.15, EtOH).

(2R)-3-methyl-2-(piperazin-2-yl)butan-2-ol

Platinum oxide (4.673 g, 20.58 mmol) was placed in a Parr bottle.Methanol (140 mL) was added under an atmosphere of nitrogen followed by(R)-(+)-3-methyl-2-(pyrazin-2-yl)butan-2-ol (17.1 g, 102.9 mmol). Thereaction mixture was shaken in a PARR hydrogenator overnight with ahydrogen pressure of 60 psi. Lc/Ms after this time indicate that thereaction was complete. The suspension was filtered through a pad ofCelite under a nitrogen atmosphere, rinsing with methanol. The crudemixture was concentrated in vacuo. The residue was re-dissolved indichloromethane, dried over MgSO4, filtered and concentrated in vacuo toafford the title compound as viscous light yellow oil (16.6 g, 3:1mixture of diastereoisomers, 93%); ¹H NMR (CDCl₃) 0.82-1.08 (9H, m),1.81 (1H, m), 2.20-3.17 (10H, m); MS ES(+) 173.0 (M+1).

4-(tert-butyldimethylsilyl)-3-chloro-2,5-difluoropyridine

A solution of diisopropylamine (78.52 g, 108.8 mL, 776.0 mmol) in THF(1.205 L) was cooled to −20° C. n-BuLi (2.5 M in hexanes) (298.4 mL of2.5 M, 746.1 mmol) was added dropwise via cannula over 30 min at such arate that the temperature was kept below −15° C. Once the addition wascomplete the cooling bath was removed and the reaction mixture wasallowed to warm to 0° C. The reaction mixture was stirred for 15 min at0° C., then re-cooled to −78° C. 3-chloro-2,5-difluoro-pyridine (95.963g, 596.9 mmol) was added dropwise via cannula over 20 min at such a ratethat the temperature was kept below −70° C. The reaction mixture wasstirred for 45 min at −78° C. during which time the solution turnedorange red. A solution of tert-butyl-chloro-dimethyl-silane (117.0 g,776.0 mmol) in THF (133.9 mL) was added via cannula at such a rate tomaintain the reaction temperature below −70° C. The reaction mixture wasstirred at −78° C. for 90 minutes, during which time the solution turnedto a deep red colour. Lc/Ms after this time indicated that the reactionwas complete. A saturated ammonium chloride solution (300 ml) was thenadded and mixture was allowed to warm to ambient. The reaction mixturewas diluted with water (200 ml) and saturated aqueous sodium bicarbonate(500 ml) and extracted with ethyl acetate (1.5 L then 500 ml). Thecombined organics were washed with brine (400 ml), dried over magnesiumsulfate, filtered and concentrated in vacuo to an oil (187 g). The crudewas purified by flash chromatography (CombiFlash Companion XL, 1.5 kgcolumn, 0.5-10% ethyl acetate in petroleum ether). This afforded thetitle compound as a pale yellow oil (113.8 g, 72%); ¹H NMR (CDCl₃) 0.46(6H, d), 0.93 (9H, s), 7.84 (1H, d); ¹⁹F NMR (decoupled) −112.8, −74.6;MS ES (+) 264.95 (M+1).

2-fluoro-N-methoxy-N-methylpyridine-3-carboxamide

2-fluoropyridine-3-carboxylic acid (100 g, 708.7 mmol) was dissolved inTHF (1.5 L). DMAP (86.58 g, 708.7 mmol) was added followed byN-methoxymethanamine hydrochloride (76.05 g, 779.6 mmol). The mixturewas cooled to 10° C. then DIPEA (100.8 g, 135.8 mL, 779.6 mmol) wasadded followed by EDC (149.5 g, 779.6 mmol) portionwise. The mixture wasstirred for 30 minutes, then warmed rapidly to ambient temp (22° C.) andstirred overnight. Lc/Ms after this time showed no acid remaining. Thereaction mixture was diluted with ethyl acetate (1 L) and water (1.5 L).The mixture was stirred for 10 minutes then the organic phase wasisolated. The aqueous was extracted with ethyl acetate (2×500 ml) andthe combined organics washed with brine, dried over magnesium sulfate,filtered and concentrated. The crude was purified through a silica plug,eluting with 50-70% ethyl acetate/petrol. This afforded the titlecompound as a pale yellow oil (81 g, 62%); ¹H NMR (CDCl₃) 3.39 (3H, s),3,68 (3H, brs), 7.29 (1H, m), 7.92 (1H, m), 8.32 (1H, m); MS ES (+)184.9 (M+1).

(4-(tert-butyldimethylsilyl)-5-chloro-3,6-difluoropyridin-2-yl)(2-fluoropyridin-3-yl)methanone

A solution of diisopropylamine (52.41 g, 72.59 mL, 517.9 mmol) in THF(967.3 mL) was cooled to −20° C. n-BuLi (2.5M in hexane) (198.5 mL of2.5 M, 496.3 mmol) was added dropwise over 15 minutes via cannulamaintaining the internal temperature below −15° C. The solution waswarmed to 0° C. then immediately re-cooled to −90° C. A solution oftert-butyl-(3-chloro-2,5-difluoro-4-pyridyl)-dimethyl-silane (113.84 g,431.6 mmol) in THF (85.35 mL) was added dropwise over 20 min via cannulaat such a rate that the internal temperature was kept below −85° C.During addition the solution first changed to bright yellow then slowlybecame an orange coloured suspension. The reaction mixture was stirredat −85° C. for 1 h. A solution of2-fluoro-N-methoxy-N-methylpyridine-3-carboxamide (91.40 g, 496.3 mmol)in THF (85.35 mL) was added dropwise over 15 minutes via cannula at sucha rate that the internal temperature was kept below −80.7° C. (ideallybelow −85° C.). The mixture turned dark orange brown after the addition.The mixture was stirred at −85° C. for 90 minutes after which time Lc/Msindicated the reaction was complete. The cooling bath was removed andthe mixture warmed to −50° C. Saturated ammonium chloride solution (400mL) was added and the mixture allowed to warm up to RT. The mixture wasdiluted with ethyl acetate (1.5 L). The aqueous phase was separated andextracted with more ethyl acetate (300 ml). The combined organics werewashed with brine (300 ml), dried over magnesium sulfate, filtered andconcentrated in vacuo to a brown oil. The crude was purified on silicagel (CombiFlash Companion XL, 1.5 kg column, 0.5-10% ethyl acetate inpetroleum ether, 0-50% ethyl acetate in petroleum ether). This gave thetitle compound as a yellow oil which solidified on standing (159.6 g,95%); ¹H NMR (CDCl₃) 0.60 (6H, d), 1.07 (9H, s), 7.47 (1H, m), 8.33 (1H,m), 8.52 (1H, m); ¹⁹F NMR (decoupled) −107.2, −72.4, −61.8; MS ES (+)387.04 (M+1).

3-(4-(tert-butyldimethylsilyl)-5-chloro-3,6-difluoropyridin-2-yl)-1H-pyrazolo[3,4-b]pyridine

[4-[tert-butyl(dimethyl)silyl]-5-chloro-3,6-difluoro-2-pyridyl]-(2-fluoro-3-pyridyl)methanone(294.86 g, 762.2 mmol) was dissolved in dioxane (2.212 L) and calciumcarbonate (152.5 g, 1.524 mol) was added. The reaction was cooled in anice bath and hydrazine hydrate (190.8 g, 185.4 mL, 3.811 mol) was addeddropwise over 15 minutes via cannula to the yellow suspension keepingthe internal temperature below 10° C. During addition, the internaltemperature dropped to 7.1° C. and the colour changed from light yellowto mango orange. The resulting mixture was allowed to warm to ambientovernight. Lc/Ms after this time indicated the reaction was complete.The reaction mixture was filtered through a pad of Celite, washingcopiously with dichloromethane (ca 2.5 L). Water (400 ml) was added tothe filtrate followed by a saturated solution of sodium bicarbonate (400mL). The organic phase separated and washed again with a saturatedaqueous sodium bicarbonate (350 mL). The combined aqueous layers wereback-extracted with dichloromethane (2×400 ml). The combined organicswere dried over magnesium sulfate, filtered and concentrated in vacuo.The resulting solid was slurried in petroleum ether, filtered and dried(81 g). The Celite was further washed with dichloromethane/methanol 7:1(3×800 ml) and with 4 L of dichloromethane/methanol 4:1 until noremaining compound was observed on TLC. The filtrate was concentrated invacuo. The residual solid was triturated with Petroleum ether asdescribed above (185 g). The solids were combined and dried at 40° C.under vacuum. This gave the title compound as an off-white solid (260.5g, 89%); ¹H NMR (CDCl₃) 0.63 (6H, d), 1.07 (9H, s), 7.44 (1H, m), 8.76(1H, m), 9.07 (1H, m), 13.31 (1H, brs); ¹⁹F NMR (decoupled) −104.7,−73.8; MS ES (+) 381.1 (M+1).

3-(4-(tert-butyldimethylsilyl)-5-chloro-3-fluoro-6-((S)-3-((R)-3-methyl-2-(trimethylsilyloxy)butan-2-yl)piperazin-1-yl)pyridin-2-yl)-1H-pyrazolo[3,4-b]pyridine

3-(4-(tert-butyldimethylsilyl)-5-chloro-3,6-difluoropyridin-2-yl)-1H-pyrazolo[3,4-b]pyridine(132.7 g, 348 mmol) was dissolved in anhydrous THF (265 mL).Imidazol-1-yl(trimethyl)silane (254 mL) was added followed by(2R)-3-methyl-2-(piperazin-2-yl)butan-2-ol (90 g, 522 mmol). The mixturewas divided into four glass pressure vessel's and the sealed containersheated and stirred at 95° C. for 48 hours. The crude mixtures werecooled, combined and diluted with ethyl acetate (1.7 L). The solutionwas washed with saturated sodium bicarbonate (2×500 ml). The combinedaqueous layers were back-extracted with ethyl acetate (500 ml). Thecombined organics were washed with brine (500 ml), dried over MgSO4,filtered and concentrated in vacuo to give a viscous brown oil. Thediastereoisomers were separated on silica gel (10-100% ethyl acetatecontaining 2% triethylamine in petroleum ether). This gave the required(S,R)-diastereoisomer as a yellow foam (125.3 g, 59%); ¹H NMR (CDCl₃)0.01 (9H, s), 0.42 (6H, s), 0.78-0.89 (15H, m), 1.09-1.13 (3H, m), 1.74(1H, m), 2.61 (1H, m), 2.80-2.99 (4H, m), 3.12 (1H, m), 3.45 (1H, d),3.69 (1H, d), 7.12 (1H, m), 8.55 (1H, m), 8.81 (1H, m), 12.75 (1H, brs);¹⁹F NMR (decoupled) −108.9; MS ES 605.2 (M+). The (R,R)-diastereoisomerwas isolated as a yellow foam (27.5 g, 13%); ¹H NMR (CDCl₃) 0.00 (9H,s), 0.38 (6H, m), 0.76-0.80 (15H, m), 0.85 (3H, s), 1.85 (1H, m), 2.50(1H, m), 2.90-3.13 (4H, m), 3.42 (1H, m), 3.56 (1H, m), 3.91 (1H, m),7.08 (1H, m), 8.51 (1H, m), 8.76 (1H, m), 12.70 (1H, brs); ¹⁹F NMR(decoupled) −109.0; MS ES 605.2 (M+).

(R)-2-((S)-4-(3-chloro-5-fluoro-6-(1H-pyrazolo[3,4-b]pyridin-3-yl)pyridin-2-yl)piperazin-2-yl)-3-methylbutan-2-ol

3-(4-(tert-butyldimethylsilyl)-5-chloro-3-fluoro-6-((S)-3-((R)-3-methyl-2-(trimethylsilyloxy)butan-2-yl)piperazin-1-yl)pyridin-2-yl)-1H-pyrazolo[3,4-b]pyridine(125 g, 206.5 mmol) was dissolved in THF (450 mL) at ambienttemperature. Tetrabutylammonium fluoride (433.6 mL of 1 M THF solution,433.6 mmol) was added dropwise via cannula to the yellow solution. Theinternal temperature increased from 21.3° C. to 27.7° C. The reactionwas stirred for 48 hours. The mixture was then diluted with ethylacetate (1.5 L) and washed with a 1:1 mixture of saturated aqueoussodium bicarbonate and water (3×600 ml). The combined aqueous phaseswere back-extracted with ethyl acetate (2×600 ml). The combined organicphases were then washed with brine (600 ml), dried over MgSO4, filteredand partially concentrated in vacuo until the title product began toprecipitate. A few drops of methanol were added and the mixture stirredat ambient temperature overnight. The product was then was filtered,rinsed with a minimum of ethyl acetate and petroleum ether to give anoff-white solid which was dried under vacuum overnight (40° C. and 1.5mbar). This afforded the title compound as a white solid (73.1 g, 84%);mp (DSC) onset 215.3° C., peak 218.1° C.; ¹H NMR (400.0 MHz, DMSO) 0.91(d, 3H), 0.94 (d, 3H), 1.10 (s, 3H), 1.83 (b quint, 2H), 2.72 (t, 1H),2.80-2.94 (m, 3H), 3.14-3.17 (m, 1H), 3.62 (d, 1H), 3.88 (d, 1H), 4.14(s, 1H), 7.37 (dd, 1H), 8.16 (d, 1H), 8.65 (dd, 1H), 8.86 (dd, 1H),14.12 (bs, 1H); ¹⁹F NMR (decoupled) −127.9; MS ES(+) 419.1 (M+1); ee>98%(Minigram SFC, ODH 250×10 mm, 20% MeOH (0.1% TEA), 5 mg/ml.

Example 3 Compound 366-((R)-3-((S)-1-amino-2-methylpropyl)pyrrolidin-1-yl)-2-chloro-5-fluoronicotinonitrile

A mixture of 2-methyl-1-[(3S)-pyrrolidin-3-yl]propan-1-amine (310 mg,1.98 mmol), 2,6-dichloro-5-fluoro-pyridine-3-carbonitrile (378.7 mg,1.98 mmol) and DIPEA (512.6 mg, 690.8 μL, 3.97 mmol) in NMP (3 mL) washeated at 130° C. for 20 minutes under microwave conditions. After thistime, the reaction mixture was allowed to cool to ambient temperatureand diluted with EtOAc, water and saturated aqueous NaHCO₃. The organiclayer was separated and washed with brine, dried (Na₂SO₄), filtered andconcentrated in vacuo. The crude mixture was purified by columnchromatography (ISCO Companion™, 40 g column, eluting with MeOH/DCM) togive the sub title compound as an oil (506.3 mg, 86% Yield).

6-(3S)-3-(1-amino-2-methylpropyl)pyrrolidin-1-yl)-5-fluoro-2-(1-trityl-1H-pyrazolo[3,4-b]pyridin-3-yl)nicotinonitrile

3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-trityl-pyrazolo[5,4-b]pyridine(1.45 g, 2.98 mmol),6-[(3S)-3-(1-amino-2-methyl-propyl)pyrrolidin-1-yl]-2-chloro-5-fluoro-pyridine-3-carbonitrile(506 mg, 1.71 mmol), K₃PO₄ (1.45 g, 6.82 mmol) and Pd(PPh₃)₄ (98.51 mg,0.09 mmol) were suspended in dioxane (18 mL) and water (1.8 mL) andheated at 100° C. for 2 hours. After this time, the reaction mixture wascooled to ambient and diluted with EtOAc, water and saturated aqueousNaHCO₃ and filtered through a celite pad. The organic layer wasseparated, washed with saturated aqueous brine, dried (Na₂SO₄), filteredand concentrated in vacuo. The crude mixture was purified by columnchromatography (ISCO Companion™, 40 g column, eluting withNH₄OH:MeOH/DCM) to give the sub title compound as an oil (762.1 mg, 72%Yield).

6-(3S)-3-(1-amino-2-methylpropyl)pyrrolidin-1-yl)-5-fluoro-2-(1H-pyrazolo[3,4-b]pyridin-3-yl)nicotinonitrile

A solution of2-[(3S)-3-(1-amino-2-methyl-propyl)pyrrolidin-1-yl]-5-fluoro-6-(1-tritylpyrazolo[3,4-b]pyridin-3-yl)pyridine-3-carbonitrile(762 mg, 1.27 mmol) in DCM (20 mL) was cooled at 0° C. and treated withtriethylsilane (142.6 mg, 195.9 μL, 1.23 mmol) and TFA (2.80 g, 1.89 mL,24.52 mmol). The reaction mixture was allowed to warm to ambienttemperature over 18 hours. After this time the reaction mixture wasconcentrated in vacuo. The residue was purified by reverse phasepreparative HPLC [Waters Sunfire C18, 5 μM, 100 Å column, gradient10%-95% B (solvent A: 0.05% TFA in water; solvent B: CH₃CN) over 16minutes at 25 mL/min] The fractions for peak 1 were collected andfreeze-dried to give the mono-TFA salt of the title compound as a solid(58.8 mg, 19.4% Yield). H¹ NMR (400.0 MHz, DMSO) δ 14.19 (s, 1H), 8.72(dd, J=1.3, 8.1 Hz, 1H), 8.62 (dd, J=1.5, 4.5 Hz, 1H), 8.03 (s, 1H),7.88 (d, J=3.0 Hz, 3H), 7.32 (dd, J=4.5, 8.1 Hz, 1H), 4.05-4.00 (m, 2H),3.75-3.11 (m, 3H), 2.60-2.50 (m, 1H), 2.26-2.18 (m, 1H), 2.01-1.74 (m,2H), 1.06 (d, J=6.9 Hz, 3H) and 1.00 (d, J=7.0 Hz, 3H) ppm; MS (ES+)380.0. The fractions for peak 2 were collected and freeze-dried to givethe mono-TFA salt of the isomer of the title compound as a solid (47.0mg, 15.2% Yield). H¹ NMR (400.0 MHz, DMSO) δ 14.18 (s, 1H), 8.74 (dd,J=1.3, 8.1 Hz, 1H), 8.62 (dd, J=1.5, 4.5 Hz, 1H), 8.04 (s, 1H), 8.00 (s,3H), 7.34 (dd, J=4.5, 8.2 Hz, 1H), 4.13-4.07 (m, 1H), 4.00-3.92 (m, 1H),3.81-3.71 (m, 1H), 3.59-3.54 (m, 1H), 3.20-3.09 (m, 1H), 2.59-2.50 (m,1H), 2.21-1.99 (m, 2H), 1.89-1.75 (m, 1H), 1.07 (d, J=7.0 Hz, 3H) and0.97 (d, J=7.0 Hz, 3H) ppm; MS (ES+) 380.0.

A 70:30 mixture of (R,S):(R,R)-piperazine b/c was resolved in five stepsby first protecting the unhindered nitrogen with Boc2O to afford amixture of (R,S):(R,R)—N-Boc piperazine d/e. This mixture was treatedwith aqueous HCHO to afford a mixture of (R,S):(R,R)—N-Boc-oxazolidinef/g. The diastereomeric mixture was separated via SiO2 columnchromatography. The pure diastereomers were sequentially deprotectedwith TfOH to form either (R,S)-oxazolidine h or (R,R)-oxazolidine andthen heating with NH2OH.HCl [or by heating the bis-(triflic acid) saltsof h or (R,R)-oxazolidine] to afford (R,S)-i or (R,R)-piperazines,respectively.

A reaction flask is equipped with a magnetic stirrer, a thermocouple,heating mantle, and a N₂ inlet. j is charged with THF to a flask. Thesuspension is cooled at 0-10° C. 1.0 M TBAF in THF (65.6 mL; 66 mmol;1.0 equiv.) is added over 5 min. The resultant solution is allowed towarm to RT. The reaction solution is diluted with water (250 mL; 10 V)which results in a thick, white suspension. After aging for 1 h, thesolid is collected by filtration (paper). The filter-cake is washed withwater (3×50 mL), EtOH (2×50 mL), and air-dry to afford k as a fine,yellow-tinted powder.

A reaction flask is equipped with a magnetic stirrer, a thermocouple,heating mantle, and a N₂ inlet. k is charged to the flask. A solution ofi is added (2.33 g; 13.5 mmol; 1.5 equiv.) in NMP (19.2 mL; 8 V). Theresultant yellow suspension is heated at 90° C. for 10 h. After coolingto RT, the resultant suspension is diluted with water (96 mL; 40 V) and1.0 M NaOH (9.9 mL; 9.9 mmol; 1.1 equiv.). The suspension is aged for 30min. The solid is collected by filtration (slow filtration). Wash thefilter-cake with water (2×60 mL) and air-dry briefly. Transfer thewet-cake to a suitable flask and remove the remaining water by ACNdilution/concentrations (2×200 mL). Fresh ACN is added (60 mL) andwarmed at 80° C. with agitation. The suspension is cololed to RT and thesolid is collected by filtration. The filter-cake is washed with ACN(2×10 mL) and air-dried to afford 9 as a fine, white powder.

HPLC: 98.7% AUC (1.3% single impurity)

¹H NMR (DMSO-d₆): conforms to structure; residual ACN.

Crystal Structure of Compound 9

Diffraction data were acquired on a Bruker Apex II diffractometerequipped with a sealed tube Cu Kα source and an Apex II CCD detector.

The structure was solved and refined using the SHELX program (Sheldrick,G. M., Acta Cryst., (2008) A64, 112-122).

Based on systematic absences and intensity statistics, the structure wassolved and refined in the orthorhombic unit cell and acentric P2₁2₁2space group.

The absolute configuration was determined reliably with −0.005(0.014)Flack's absolute configuration factor. The value is very close to 0 witha small standard deviation. This indicates the absolute configuration isdetermined reliably.

There is one molecule in the asymmetric unit.

There exists a zigzag one-dimensional infinite chain with R 2,2(9) graphset for the hydrogen bonding synthon structure. There is no hydrogenbonding between such chains, only van der Waals interaction.

Comparison of the simulated and calculated patterns indicates thestructure represents the batch.

The structure is fully ordered with a low R factor of 2.3%. We give ascore of A for the quality of the structure.

EXPERIMENTAL

Dissolved 20 mg of compound 9 in lmL methanol, 4 mL ethanol, addedseeds, heated at 80° C. in a sealed vial. Needle shape crystals wereobtained over night.

A colorless blade shape crystal with dimensions of 0.30×0.10×0.05 mm³was selected, mounted on a MicroMount and centered on a Bruker APEX IIdiffractometer (V011510). Three batches of 40 frames separated inreciprocal space were obtained to provide an orientation matrix andinitial cell parameters. The final cell parameters were collected andrefined was completed based on the full data set.

A diffraction data set of reciprocal space was obtained to a resolutionof 106° 2θ angle using 0.5° steps with exposure times 10 s each framefor low angle frames and 60 s each frame for high angle frames. Datawere collected at 100 (2) K temperature. Integration of intensities andrefinement of cell parameters were conducted using the APEXII software.

The powder X-ray diffraction data were collected on a Bruker D8 Discoverdiffractometer with Cu Kα radiation and a Highstar detector.

Refinement

The structure is fully ordered. Hydrogen atoms were refined using theriding model.

Computing Details

Data collection: Apex II; cell refinement: Apex II; data reduction: ApexII; program(s) used to solve structure: SHELXS97 (Sheldrick, 1990);program(s) used to refine structure: SHELXL97 (Sheldrick, 1997);molecular graphics: Mercury; software used to prepare material forpublication: publCIF.

Crystal Data

Crystal data

C₂₀H₂₄ClFN₆O D_(x) = 1.340 Mg m⁻³ M_(r) = 418.90 Cu Kα radiation, λ =1.54178 Å Orthorhombic, P2₁2₁2 Cell parameters from 5449 reflections a =15.0138 (8) Å θ = 3.4-52.8° b = 24.9923 (14) Å μ = 1.91 mm⁻¹ c = 5.5319(3) Å T = 100 K V = 2075.7 (2) Å³ Blade, colorless Z = 4 0.30 × 0.10 ×0.05 mm F(000) = 880

Data collection

Bruker APEX-II CCD 2283 reflections with I > 2σ(I) diffractometerRadiation source: fine-focus R_(int) = 0.030 sealed tube graphiteθ_(max) = 52.9°, θ_(min) = 3.4° φ and ω scans h = −15→13 8508 measuredreflections k = −25→25 2382 independent reflections l = −5→5

Refinement

Refinement on F² Hydrogen site location: inferred from neighbouringsites Least-squares H atoms treated by a mixture of matrix: fullindependent and constrained refinement R[F² > 2σ(F²)] = 0.023 w =1/[σ²(F_(o) ²) + (0.0336P)² + 0.3794P] where P = (F_(o) ² + 2F_(c) ²)/3wR(F²) = 0.059 (Δ/σ)_(max) = 0.004 S = 1.04 Δ

_(max) = 0.12 e Å⁻³ 2382 reflections Δ

_(min) = −0.11 e Å⁻³ 359 parameters Extinction correction: SHELXL, Fc* =kFc[1 + 0.001 × Fc²λ³/sin(2θ)]^(−1/4) 0 restraints Extinctioncoefficient: 0.00037 (14) Primary atom site location: Absolutestructure: Flack H D (1983), structure-invariant Acta Cryst. A39,876-881 direct methods Secondary atom site location: Flack parameter:−0.005 (14) difference Fourier mapSpecial Details

Geometry.

All esds (except the esd in the dihedral angle between two l.s. planes)are estimated using the full covariance matrix. The cell esds are takeninto account individually in the estimation of esds in distances, anglesand torsion angles; correlations between esds in cell parameters areonly used when they are defined by crystal symmetry. An approximate(isotropic) treatment of cell esds is used for estimating esds involvingl.s. planes.

Refinement.

Refinement of F² against ALL reflections. The weighted R-factor wR andgoodness of fit S are based on F², conventional R-factors R are based onF, with F set to zero for negative F². The threshold expression of F²>2sigma(F²) is used only for calculating R-factors (gt) etc. and is notrelevant to the choice of reflections for refinement. R-factors based onF² are statistically about twice as large as those based on F, andR-factors based on ALL data will be even larger.

TABLE A Fractional atomic coordinates and isotropic or equivalentisotropic displacement parameters (Å²) x y z U_(iso)*/U_(eq) Cl1 1.04404(4) 0.58029 (2) 0.34143 (10) 0.03616 (19) F1 1.20454 (8) 0.66248 (5)1.0451 (2) 0.0366 (3) O1 0.68641 (11) 0.57143 (6) 0.4801 (3) 0.0272 (4)N6 0.74240 (12) 0.67589 (7) 0.2358 (4) 0.0249 (5) N1 0.99956 (11)0.70617 (7) 0.7671 (3) 0.0247 (4) N2 1.14128 (12) 0.75283 (7) 1.2551 (3)0.0285 (5) N5 0.91316 (11) 0.67077 (7) 0.4524 (3) 0.0237 (4) C1 1.07209(13) 0.70611 (9) 0.9140 (4) 0.0239 (5) C2 0.86382 (16) 0.72146 (9)0.4486 (4) 0.0254 (5) C3 0.79085 (14) 0.62439 (9) 0.2451 (4) 0.0236 (5)N3 1.12675 (12) 0.79799 (8) 1.3849 (4) 0.0282 (5) C4 0.98751 (13)0.66811 (9) 0.6028 (4) 0.0233 (5) C5 1.05189 (14) 0.62702 (8) 0.5716 (4)0.0271 (5) C6 0.84970 (15) 0.62586 (9) 0.4687 (4) 0.0246 (5) C7 0.72662(13) 0.57558 (9) 0.2458 (4) 0.0256 (5) C8 1.07779 (13) 0.75034 (8)1.0867 (4) 0.0240 (5) N4 1.02367 (11) 0.87124 (7) 1.3936 (3) 0.0288 (5)C9 1.05411 (14) 0.82485 (9) 1.3027 (4) 0.0253 (5) C10 0.78037 (15)0.52250 (9) 0.2120 (4) 0.0272 (6) C11 0.94497 (14) 0.81731 (9) 0.9897(4) 0.0285 (6) C12 1.13329 (13) 0.66481 (9) 0.8951 (4) 0.0270 (5) C131.12472 (15) 0.62587 (10) 0.7230 (4) 0.0293 (6) C14 0.65471 (15) 0.58205(11) 0.0558 (4) 0.0286 (5) C15 0.95206 (15) 0.88963 (9) 1.2770 (4)0.0327 (6) C16 1.01947 (13) 0.79594 (9) 1.1065 (4) 0.0247 (5) C170.80485 (15) 0.72167 (9) 0.2276 (4) 0.0269 (6) C18 0.72073 (19) 0.47349(12) 0.2330 (7) 0.0439 (7) C19 0.91192 (16) 0.86462 (9) 1.0796 (4)0.0324 (6) C20 0.83300 (19) 0.52023 (12) −0.0247 (5) 0.0398 (7) H10.8284 (13) 0.6202 (8) 0.108 (4) 0.014 (5)* H2 0.8286 (13) 0.7276 (8)0.603 (4) 0.021 (6)* H3 0.8822 (12) 0.5928 (8) 0.486 (3) 0.010 (5)* H40.6810 (15) 0.5897 (9) −0.100 (5) 0.035 (6)* H5 0.6204 (14) 0.5493 (9)0.044 (4) 0.030 (6)* H6 0.7118 (14) 0.6754 (9) 0.102 (4) 0.023 (6)* H70.7682 (13) 0.7558 (8) 0.218 (3) 0.014 (5)* H8 0.9025 (14) 0.7517 (9)0.432 (4) 0.023 (6)* H9 0.8398 (14) 0.7190 (8) 0.080 (4) 0.017 (5)* H100.8252 (14) 0.5220 (7) 0.353 (4) 0.017 (5)* H11 0.6125 (15) 0.6120 (9)0.096 (4) 0.032 (6)* H12 0.8104 (14) 0.6294 (8) 0.618 (4) 0.022 (5)* H130.6865 (19) 0.4754 (10) 0.378 (5) 0.052 (8)* H14 1.1690 (15) 0.5983 (9)0.697 (4) 0.032 (6)* H15 0.684 (2) 0.4701 (12) 0.097 (6) 0.064 (9)* H160.9303 (14) 0.9226 (9) 1.342 (4) 0.033 (6)* H17 0.9200 (14) 0.8009 (8)0.854 (4) 0.027 (6)* H18 0.882 (2) 0.5493 (13) −0.039 (6) 0.082 (10)*H19 0.8631 (15) 0.4848 (10) −0.038 (4) 0.033 (6)* H20 0.6366 (19) 0.5853(12) 0.489 (6) 0.061 (10)* H24 0.7569 (15) 0.4408 (10) 0.243 (4) 0.032(6)* H22 0.8643 (15) 0.8801 (9) 1.003 (4) 0.034 (6)* H21 1.1660 (19)0.8064 (11) 1.524 (5) 0.058 (8)* H23 0.7900 (17) 0.5223 (9) −0.164 (5)0.047 (7)*

TABLE B Atomic displacement parameters (Å²) U¹¹ U²² U³³ U¹² U¹³ U²³ Cl10.0319 (3) 0.0383 (3) 0.0383 (3) 0.0041 (3) 0.0019 (3) −0.0099 (3) F10.0253 (7) 0.0380 (8) 0.0465 (8) 0.0030 (6) −0.0099 (6) −0.0001 (7) O10.0256 (9) 0.0357 (10) 0.0202 (8) 0.0003 (8) 0.0018 (7) 0.0010 (7) N60.0251 (11) 0.0279 (12) 0.0216 (11) 0.0006 (9) −0.0032 (9) 0.0011 (9) N10.0217 (10) 0.0266 (11) 0.0256 (10) −0.0037 (8) 0.0005 (9) 0.0018 (9) N20.0254 (11) 0.0312 (11) 0.0290 (11) −0.0024 (8) −0.0017 (9) 0.0020 (10)N5 0.0228 (10) 0.0239 (10) 0.0244 (9) 0.0001 (8) −0.0006 (8) 0.0010 (8)C1 0.0201 (12) 0.0271 (13) 0.0244 (12) −0.0027 (9) 0.0001 (10) 0.0042(11) C2 0.0254 (13) 0.0249 (14) 0.0258 (14) −0.0002 (11) 0.0006 (11)0.0011 (11) C3 0.0253 (12) 0.0298 (14) 0.0156 (12) 0.0053 (10) 0.0034(11) 0.0014 (10) N3 0.0255 (11) 0.0323 (11) 0.0268 (11) −0.0009 (9)−0.0031 (9) −0.0005 (10) C4 0.0198 (12) 0.0273 (12) 0.0226 (12) −0.0028(10) 0.0027 (10) 0.0047 (11) C5 0.0244 (12) 0.0263 (12) 0.0305 (13)−0.0013 (10) 0.0039 (11) 0.0008 (10) C6 0.0242 (13) 0.0255 (14) 0.0240(13) 0.0013 (10) −0.0003 (11) 0.0017 (11) C7 0.0259 (12) 0.0297 (13)0.0214 (12) 0.0014 (10) 0.0014 (9) −0.0016 (10) C8 0.0187 (12) 0.0292(13) 0.0239 (13) −0.0043 (10) −0.0004 (10) 0.0050 (10) N4 0.0225 (11)0.0351 (12) 0.0288 (10) −0.0035 (9) 0.0044 (8) −0.0029 (9) C9 0.0219(12) 0.0301 (13) 0.0240 (12) −0.0043 (11) 0.0036 (11) 0.0032 (11) C100.0254 (13) 0.0306 (14) 0.0257 (13) 0.0008 (10) −0.0045 (11) −0.0015(11) C11 0.0237 (13) 0.0323 (14) 0.0296 (13) −0.0060 (11) −0.0028 (12)0.0009 (11) C12 0.0189 (12) 0.0321 (14) 0.0300 (13) −0.0038 (11) −0.0062(11) 0.0047 (12) C13 0.0234 (13) 0.0290 (14) 0.0354 (14) 0.0025 (11)0.0031 (12) 0.0039 (12) C14 0.0265 (13) 0.0340 (15) 0.0252 (14) 0.0008(13) −0.0043 (11) −0.0036 (12) C15 0.0279 (14) 0.0305 (15) 0.0397 (15)0.0003 (12) 0.0052 (13) −0.0042 (12) C16 0.0196 (13) 0.0291 (13) 0.0254(12) −0.0047 (10) 0.0019 (10) 0.0028 (11) C17 0.0275 (14) 0.0287 (15)0.0244 (14) 0.0036 (11) 0.0025 (12) 0.0041 (11) C18 0.0374 (17) 0.0312(17) 0.063 (2) 0.0016 (13) −0.0040 (17) −0.0067 (15) C19 0.0235 (13)0.0330 (15) 0.0408 (15) 0.0017 (11) −0.0040 (12) 0.0010 (12) C20 0.0493(17) 0.0403 (18) 0.0299 (15) 0.0156 (14) 0.0038 (14) −0.0062 (13)

TABLE C Geometric parameters (Å, °) Cl1—C5 1.732 (2) C3—C7 1.555 (3)F1—C12 1.355 (2) N3—C9 1.359 (3) O1—C7 1.434 (2) C4—C5 1.421 (3) N6—C31.479 (3) C5—C13 1.377 (3) N6—C17 1.480 (3) C7—C14 1.515 (3) N1—C4 1.328(3) C7—C10 1.564 (3) N1—C1 1.359 (3) C8—C16 1.441 (3) N2—C8 1.334 (3)N4—C15 1.335 (3) N2—N3 1.355 (3) N4—C9 1.344 (3) N5—C4 1.394 (3) C9—C161.404 (3) N5—C2 1.468 (3) C10—C18 1.522 (3) N5—C6 1.475 (3) C10—C201.530 (3) C1—C12 1.386 (3) C11—C19 1.375 (3) C1—C8 1.463 (3) C11—C161.398 (3) C2—C17 1.509 (3) C12—C13 1.368 (3) C3—C6 1.520 (3) C15—C191.395 (3) C3—N6—C17 111.23 (17) O1—C7—C10 105.31 (17) C4—N1—C1 121.17(17) C14—C7—C10 112.04 (18) C8—N2—N3 107.09 (17) C3—C7—C10 110.19 (17)C4—N5—C2 116.98 (17) N2—C8—C16 110.11 (18) C4—N5—C6 116.39 (16) N2—C8—C1122.21 (18) C2—N5—C6 109.38 (17) C16—C8—C1 127.68 (18) N1—C1—C12 119.15(19) C15—N4—C9 112.96 (19) N1—C1—C8 115.88 (18) N4—C9—N3 125.0 (2)C12—C1—C8 124.96 (19) N4—C9—C16 127.4 (2) N5—C2—C17 108.08 (18)N3—C9—C16 107.57 (19) N6—C3—C6 107.03 (18) C18—C10—C20 109.8 (2)N6—C3—C7 112.19 (17) C18—C10—C7 111.72 (19) C6—C3—C7 112.17 (17)C20—C10—C7 113.59 (19) N2—N3—C9 111.29 (18) C19—C11—C16 116.7 (2)N1—C4—N5 118.91 (18) F1—C12—C13 118.04 (19) N1—C4—C5 120.54 (18)F1—C12—C1 120.60 (19) N5—C4—C5 120.45 (18) C13—C12—C1 121.3 (2)C13—C5—C4 118.8 (2) C12—C13—C5 118.9 (2) C13—C5—Cl1 119.12 (17)N4—C15—C19 124.9 (2) C4—C5—Cl1 122.03 (16) C11—C16—C9 117.2 (2) N5—C6—C3110.12 (18) C11—C16—C8 138.8 (2) O1—C7—C14 109.56 (17) C9—C16—C8 103.93(18) O1—C7—C3 108.67 (16) N6—C17—C2 110.14 (18) C14—C7—C3 110.88 (18)C11—C19—C15 120.8 (2)

The following compounds can be synthesized, in general, based on asimilar route to that outlined in Example 1, 2 and 3, Compounds 2-8,10-35 and 37-45.

Similar methods are described in PCT Application No. PCT/US2009/051437filed Jul. 22, 2009 entitled “TRI-CYCLIC PYRAZOLOPYRIDINE KINASEINHIBITORS” the entire contents of which are incorporated herein byreference.

Table 2 depicts data for certain exemplary compounds made in general bya similar route to that outlined in the above Examples.

TABLE 2 M + 1 RT No. (obs) (min) 1H-NMR 1 403 7.47 1H NMR (400 MHz,DMSO) d 13.99 (s, 1H), 8.77 (dd, J = 8.1, 1.4 Hz, 1H), 8.60 (dd, J =4.4, 1.5 Hz, 1H), 7.92 (dd, J = 12.1, 9.8 Hz, 1H), 7.30 (dd, J = 8.1,4.5 Hz, 1H), 4.14 (s, 1H), 4.04 (d, J = 7.9 Hz, 1H), 3.77 (d, J = 8.6Hz, 1H), 3.09 (d, J = 8.9 Hz, 1H), 2.84 (d, J = 9.1 Hz, 2H), 2.74 (d, J= 7.4 Hz, 2H), 1.81 (dt, J = 13.5, 6.8 Hz, 2H), 1.05 (s, 3H), 0.87 (dd,J = 23.8, 6.7 Hz, 6H). 2 401.2 2.36 1H (DMSO) d 0.15 (d, 1H), 0.20 (d,1H), 0.89 (s, 3H), 0.96 (s, 3H), 1.53 (bs, 1H, NH), 2.24-2.31 (m, 1H),2.64-2.72 (m, 2H), 2.79 (t, 1H), 2.91 (d, 1H), 3.58-3.63 (m, 2H), 4.70(s, 1H, OH), 7.10 (dd, 1H), 7.72 (dd, 1H), 8.40 (dd, 1H), 8.55 (1H, dd),13.80 (bs, 1H), NH) ppm 3 403 7.67 4 401 7.42 5 401 7.52 6 389 7.17 1HNMR (CD3OD) 1.05 (3H, t), 1.28-1.32 (4H, m), 1.60-1.85 (2H, m), 2.98(3H, m), 3.10-3.15 (2H, m), 4.05-4.10 (1H, m), 4.30-4.35 (1H, m),7.40-7.45 (1H, dd), 7.70-7.80 (1H, dd), 8.70 (1H, d), 9.00 (1H, d). 7389 1.68 (CD3OD) 1.01 (6H, d), 1.25-1.35 (1H, m), 1.85-1.95 (1H, m),1.85-1.90 (1H, m), 2.95-3.10 (2H, m), 3.20 (1H, M), 3.70 (1H, s),3.92-3.97 (1H, m), 4.20-4.25 (1H, d), 7.33-7.36 (1H, dd), 7.59-7.64 (1H,m), 8.57-8.59 (1H, d), 8.87-8.89 (1H, d). 8 375 6.91 DMSO-d6 0.89-0.93(3H, m), 1.3-1.4 (1H, m), 1.55-1.60 (1H, m), 2.65-2.70 (2H, m) 2.80-3.00(2H, m), 3.02-3.07 (1H, m), 3.25-3.30 (1H, m), 3.50 (1H, s), 3.75-3.80(1H, m), 4.03-4.08 (1H, m), 4.59-4.61 (1H, m), 7.29-7.32 (1H, d),7.90-7.95 (1H, dd), 8.59-8.60 (1H, d), 8.76-8.79 (1H, d), 13.95-13.90(1H, s). 9 419 8.27 (CD3OD) 0.97-1.03 (6H, dd), 1.21 (3H, s), 1.93-1.94(1H, m), 2.80-2.86 (1H, t), 3.00-3.08 (2H, m), 3.22-3.25 (1H, m),3.71-3.75 (1H, d), 3.99-4.03 (1H, d), 7.32-7.35 (1H, d), 7.83-7.86 (1H,d), 8.58-8.59 (1H, d), 8.90-8.92 (1H, d). 10 (DMSO, 400 MHz) 0.83 (3H,s), 0.94 (3H, s), 1.09 (3H, s), 1.87 (1H, sept), 3.15-3.50 (5H, m), 4.18(1H, d), 4.38 (1H, d), 7.37 (1H, dd), 8.46 (1H, d), 8.66 (1H, dd), 8.71(1H, d), 14.40 (1H, br s). 11 421.2 2.35 1H (DMSO) d 1.24 (d, 3H), 1.36(d, 3H), 1.42 (d, 3H), 2.59-2.67 (m, 1H), 2.88-2.93 (m, 2H), 2.98 (dd,1H), 3.14 (d, 1H), 3.78 (d, 1H), 4.09 (d, 1H), 4.86 (s, 1H, OH), 7.25(dd, 1H), 7.92 (dd, 1H), 8.58 (dd, 1H), 8.74 (dd, 1H), 13.99 (bs, 1H,NH) ppm 12 421.2 2.31 1H (DMSO) d 1.21 (s, 3H), 1.40 (t, 6H), 1.97 (bs,1H, NH), 2.75-2.95 (m, 4H), 3.06 (d, 1H), 3.76 (d, 1H), 4.16 (d, 1H),4.77 (s, 1H, OH), 7.30 (dd, 1H), 7.92 (dd, 1H), 8.59 (dd, 1H), 8.82 (dd,1H), 13.97 (bs, 1H, NH) ppm 13 429 0.77 (d6-DMSO, 400 MHz) 1.49 (3H, s),3.15-3.20 (1H, m), 3.21-3.45 (3H, m), 3.78 (1H, brs), 4.11 (1H, d), 4.19(1H, d), 7.30 (1H, dd), 7.45 (brs), 8.08 (1H, dd), 8.62-8.64 (2H, m),8.92 (1H, brs), 9.09 (1H, brs), 14.09 (1H, brs) 14 429 0.73 (d6-DMSO,400 MHz) 1.49 (3H, s), 3.18 (1H, t), 3.27-3.33 (1H, m), 3.37-3.45 (2H,m), 3.79 (1H, t), 4.11 (1H, d), 4.19 (1H, d), 7.29 (1H, dd), 7.44 (1H,s), 8.09 (1H, dd), 8.62 (2H, m), 8.91 (1H, brs), 9.05 (1H, brs), 14.09(1H, brs) 15 443 8.35 1H NMR (CD3OD) 1.30 (3H, s), 2.30-2.60 (1H, m),2.65-2.80 (1H, m), 2.85-2.95 (2H, m), 3.00-3.10 (1H, m), 3.80-3.85 (1H,m), 4.05-4.10 (1H, m), 7.25 (1H, d), 7.50-7.55 (1H, m), 8.46-8.48 (1H,d), 8.73-8.76 (1H, d). 16 405 6.97 1H NMR (CD3OD) 1.32 (3H, s),3.20-3.25 (1H t), 3.30-3.45 (5H, masked), 3.63-3.67 (1H, m), 4.21-4.25(1H, m), 4.35-4.39 (1H, m), 7.32-7.35 (1H, dd), 7.71-7.76 (1H, dd),8.60-8.62 (1H, d), 8.74-8.76 (1H, d). 17 405 6.89 H NMR (400.0 MHz,DMSO) d 13.99 (s, 1H, NH), 8.79 (dd, J = 1.4, 8.1 Hz, 1H), 8.60 (dd, J =1.5, 4.5 Hz, 1H), 7.93 (dd, J = 9.9, 12.0 Hz, 1H), 7.30 (dd, J = 4.5,8.1 Hz, 1H), 3.98-3.92 (m, 1H), 3.80-3.77 (m, 1H), 3.77 (s, 1H, OH),3.38-3.32 (m, 1H), 3.27-3.24 (m, 4H), 3.08 (d, 1H), 2.87-2.67 (m, 4H),2.15 (bs, 1H, NH) and 1.16 (s, 3H) ppm 18 391 7.44 19 391 7.48 H NMR(400.0 MHz, DMSO) d 0.92 (t, 3H), 1.34 (septet, 1H), 1.56-1.65 (m, 1H),2.61 (t, 1H), 2.67-2.72 (m, 1H), 2.82-2.92 (m, 2H), 3.03-3.06 (m, 1H),3.26-3.34 (masked signal, 1H), 3.58 (d, 1H), 3.87 (d, 1H), 4.56 (d, 1H),7.34 (dd, 1H), 8.11 (d, 1H), 8.61 (dd, 1H), 8.81 (dd, 1H) and 14.10 (brs, 1H) ppm 20 405 1.05 1H (DMSO-d6) 0.89-0.96 (6H, dd), 1.85-1.90 (1H,m), 2.60-2.65 (1H, m), 2.78-2.82 (1H, m), 2.88-2.93 (1H, m), 3.10-3.15(1H, m), 3.20 (1H, m), 3.65-3.67 (1H, m), 3.92-3.95 (1H, m), 4.54-4.54(1H, m), 7.40-7.43 (1H, m), 8.14-8.16 (1H, d), 8.70 (1H, s), 8.75 (1H,d), 21 405 1.06 1H (DMSO-d6) 0.91-0.95 (6H, m), 1.80-1.85 (1H, m),2.70-2.75 (1H, m), 2.85-3.00 (2H, m), 3.10-3.20 (2H, m), 3.65-3.67 (2H,m), 4.65-4.68 (1H, m), 7.37-7.40 (1H, m), 8.15-8.18 (1H, d), 8.64-8.66(1H, d), 8.81 (1H, d), 14.1-1.2 (1H, br s) 22 373 2.42 (d6-DMSO, 400MHz) 0.99 (3H, d), 1.06 (3H, d), 1.75-1.80 (1H, m), 1.93-1.96 (1H, m),2.16-2.21 (1H, m), 3.07-3.17 (2H, m), 3.43 (1H, t), 3.62-3.66 (1H, m),3.83 (2H, brs), 7.28 (1H, dd), 7.81-7.88 (4H, m), 8.59 (1H, dd), 8.76(1H, d), 13.96 (1H, brs) 23 389.17 2.54 H NMR (400.0 MHz, DMSO) d 0.99(d, 3H), 1.06 (d, 3H), 1.71-1.79 (m, 1H), 1.91-1.99 (m, 1H), 2.20 (qn,1H), 3.13-3.20 (m, 1H), 3.59 (t, 2H), 3.73-3.85 (m, 3H), 7.29 (dd, 1H),7.89 (br s, NH3+, 3H), 7.99 (d, 1H), 8.60 (dd, 1H), 8.76 (dd, 1H) and14.07 (s, 1H) ppm 24 389.17 2.6 H NMR (400.0 MHz, DMSO) d 0.96 (d, 3H),1.06 (d, 3H), 1.79 (quintet, 1H), 2.01-2.15 (m, 2H), 3.14 (br m, 1H),3.20-3.60 (masked signal, 1H), 3.63 (t, 1H), 3.71 (t, 1H), 3.79-3.86 (m,2H), 7.32 (dd, 1H), 7.83 (s, NH3+, 3H), 8.00 (d, 1H), 8.61 (dd, 1H),8.76 (dd, 1H) and 14.07 (s, 1H) ppm 25 375.2 2.28 1H NMR (400.0 MHz,DMSO) d 14.00 (bs, 1H, NH), 8.73 (dd, J = 1.5, 8.1 Hz, 1H), 8.60 (dd, J= 1.5, 4.5 Hz, 1H), 7.93 (dd, J = 9.9, 12.1 Hz, 1H), 7.30 (dd, J = 4.5,8.1 Hz, 1H), 4.67 (d, J = 5.6 Hz, 1H), 3.82-3.74 (m, 2H), 3.35-3.30 (m,2H), 3.09-3.03 (m, 1H), 2.98-2.91 (m, 1H), 2.86-2.80 (m, 1H), 2.73-2.68(m, 2H), 2.33-2.28 (bs, 1H, NH), 1.57-1.53 (m, 1H), 1.38-1.34 (m, 1H)and 0.92 (t, J = 7.4 Hz, 3H) ppm 26 419 8.69 1H NMR (dmso-d6) 0.98 (9H,s), 2.80-3.00 (4H, m), 3.10-3.25 (2H, m), 3.60-3.64 (1H, m), 3.82-3.85(1H, m), 7.34-7.38 (1H, dd), 8.13-8.16 (1H, d), 8.64-8.66 (1H, dd),8.85-8.88 (1H, dd)., 27 403 8.23 1H NMR (CD3OD) 1.01 (9H, s), 2.95-3.20(56H, m), 3.90-3.95 (1H, d), 4.10-.4-15 (1H, d), 7.29-7.32 (1H, d), 7.59(1H, dd), 8.55-8.57 (1H, d), 8.83-8.84 (1H, d) 28 373.18 2.44 H NMR(400.0 MHz, DMSO) d 0.87 (d, 3H), 0.95 (d, 3H), 1.58-1.60 (m, 1H),1.69-1.76 (m, 1H), 2.13 - 2.28 (m, 2H), 2.46 (dd, 1H), 3.32-3.36 (maskedsignal, 1H), 3.57-3.64 (m, 1H), 3.70-3.80 (m, 2H), 7.29 (dd, 1H), 7.79(dd, 1H), 8.57 (dd, 1H) and 8.75 (dd, 1H) ppm 29 387 1.77 1H NMR (dmso)0.28-.029 (2H, s), 0.35-0.37 (1H, m), 0.39-0.40 (1H, m), 0.97-0.99 (1H,m), 2.80-2.95 (5H, m), 3.06-3.09 (1H, m), 3.77-3.79 (1H, m), 4.04-4.0(1H, m), 4.72 (1H, s), 7.28-7.31 (1H, d), 7.90-7.96 (1H, dd), 8.59-8.60(1H, d), 8.78-8.80 (1H, m), 13.90-1.40 (1H, s). 30 417 0.64 (d6-DMSO,400 MHz) 0.26-0.30 (3H, m), 0.45-0.48 (1H, m), 0.93-1.03 (1H, m), 1.11(3H, s), 2.75-2.93 (3H, m), 3.12 (1H, d), 3.63 (1H, d), 3.92 (1H, d),4.10 (1H, brs), 7.32 (1H, dd), 8.13 (1H, d), 8.62 (1H, dd), 8.85 (1H,dd), 14.10 (1H, brs) 31 403 1.91 1H NMR (DMSO-d6) 0.0-0.05 (2H, m),0.07-0.10 (1H, m), 0.20-0.25 (1H, m), 0.70-0.80 (2H, m), 2.50-2.70 (4H,m), 2.82-2.90 (1H, m), 3.30-3.40 (1H, m), 3.60-3.65 (1H, m), 7.07-7.10(1H, d), 7.85-7.88 (1H, m), 8.36-8.37 (1H, d), 8.57-8.59 (1H, d). 32 4010.51 (d6-DMSO, 400 MHz) 0.84 (3H, d), 0.94 (3H, d), 1.09 (2H, s),1.76-1.88 (1H, m), 2.89 (1H, t), 3.13-3.20 (1H, m), 3.27-3.39 (3H, m),4.08 (1H, d), 4.33 (1H, d), 5.20 (1H, brs), 7.17 (1H, d), 7.26 (1H, dd),8.33-8.40 (1H, m), 8.55-8.59 (2H, m), 8.66 (1H, dd), 11.00 (1H, brs),13.83 (1H, brs) 33 421.1 2.35 H NMR (400.0 MHz, DMSO) d 14.10 (bs, 1H,NH), 8.83 (dd, J = 1.6, 8.1 Hz, 1H), 8.62 (dd, J = 1.5, 4.5 Hz, 1H),8.12 (d, J = 9.8 Hz, 1H), 7.33 (dd, J = 4.5, 8.1 Hz, 1H), 4.57 (s, 1H),3.77 (d, J = 11.3 Hz, 1H), 3.61 (d, J = 9.0 Hz, 1H), 3.35 (d, J = 11.0Hz, 1H), 3.28-3.23 (m, 4H), 3.11-3.08 (m, 1H), 2.86-2.79 (m, 3H),2.73-2.67 (m, 1H), 2.12 (bs, 1H, NH) and 1.16 (s, 3H) ppm 34 410 0.98 HNMR (400.0 MHz, DMSO) d 14.28 (s, 1H), 8.80-8.57 (m, 3H), 8.28 (d, 1H),7.35 (dd, J = 4.5, 8.1 Hz, 1H), 4.81-4.37 (m, 2H), 3.57-3.21 (m, 5H),1.96-1.81 (m, 1H), 1.06 (d, J = 4.9 Hz, 3H), 0.93 (d, J = 6.6 Hz, 3H)and 0.67 (d, J = 6.8 Hz, 3H) ppm 35 410 0.98 H NMR (400.0 MHz, DMSO) d14.28 (s, 1H), 8.85-8.53 (m, 4H), 8.28 (s, 1H), 7.34 (dd, J = 4.5, 8.1Hz, 1H), 4.70-4.47 (m, 2H), 3.56-3.21 (m, 5H), 1.14 (s, 3H) and0.88-0.79 (m, 6H) ppm 36 380 0.59 H NMR (400.0 MHz, DMSO) d 14.19 (s,1H), 8.72 (dd, J = 1.3, 8.1 Hz, 1H), 8.62 (dd, J = 1.5, 4.5 Hz, 1H),8.03 (s, 1H), 7.88 (d, J = 3.0 Hz, 3H), 7.32 (dd, J = 4.5, 8.1 Hz, 1H),4.05-4.00 (m, 2H), 3.75-3.11 (m, 3H), 2.60-2.50 (m, 1H), 2.26-2.18 (m,1H), 2.01-1.74 (m, 2H), 1.06 (d, J = 6.9 Hz, 3H) and 1.00 (d, J = 7.0Hz, 3H) ppm 37 380 0.59 H NMR (400.0 MHz, DMSO) d 14.31 (s, 1H), 8.79(d, 1H), 8.65 (m, 1H), 8.25 (d, 1H), 7.85 (brs, 3H), 7.35 (dd, 1H),4.04-3.94 (m, 2H), 3.82-3.71 (m, 1H), 3.62-3.15 (m, 2H), 2.71-2.44 (m,1H), 2.29-2.18 (m, 1H), 2.01-1.75 (m, 2H), 1.08 (d, 3H) and 1.01 (d, 3H)ppm 38 419 0.67 (d6-DMSO, 400 MHz) 0.91 (6H, dd), 1.07 (3H, s),1.91-1.98 (1H, m), 2.63-2.71 (1H, m), 2.88-2.93 (2H, m), 3.18 (1H, d),3.63 (1H, d), 3.78 (1H, d), 4.13 (1H, s), 7.37 (1H, dd), 8.15 (1H, d),8.65 (1H, dd), 8.83 (1H, d), 14.12 (1H, brs) 39 419 1.77 40 419 2 41 4192.05 42 435 2.54 43 (DMSO, 400 MHz) 0.77 (3H, d), 0.89 (3H, d), 1.05(3H, s), 1.83 (1H, sept), 2.70-3.20 (5H, m), 4.31 (1H, d), 4.65 (1H, d),7.34 (1H, dd), 8.11 (1H, d), 8.63 (1H, d), 8.70 (1H, d), 14.22 (1H, brs). 44 415.2 2.05 H NMR (400.0 MHz, DMSO) d 14.01 (bs, 1H, NH), 8.82 (d,1H), 8.60 (dd, 1H), 7.79 (d, 1H), 7.30 (d, 1H), 5.54 (t, 1H), 4.60-4.50(m, 2H), 4.28 (bs, 1H), 3.48-3.36 (m, 2H), 3.18-3.13 (m, 1H), 2.98-2.89(m, 2H), 2.73-2.66 (m, 1H), 1.83-1.76 (m, 1H), 1.06 (s, 3H), 0.91 (d,3H) and 0.85 (d, 3H) ppm 45 435 2.23

In general, compounds of the invention, including compounds in Table 1,are effective for the inhibition of PKCtheta. Selectivity for inhibitionof PKCtheta by the compounds of the invention was tested and the resultsare shown in the following Example. The data obtained shows values forPKCtheta isoform selectivity by showing Ki potencies for PKCtheta,PKCdelta and PKCalpha.

Example 4 PKC Theta

An assay buffer solution was prepared which consisted of 100 mM HEPES(pH 7.5), 10 mM MgCl₂, 25 mM NaCl, 0.1 mM EDTA and 0.01% Brij. An enzymebuffer containing reagents to final assay concentrations of 0.00001%Triton X-100, 200 μg/mL Phosphatidylserine, 20 μg/mL Diacylglycerol, 360μM NADH, 3 mM phosphoenolpyruvate, 70 μg/mL pyruvate kinase, 24 μg/mLlactate dehydrogenase, 2 mM DTT, 100 μM substrate peptide(ERMRPRKRQGSVRRRV SEQ ID NO. 1) and 18 nM PKC theta kinase was preparedin assay buffer. To 60 μL of this enzyme buffer, in a 384 well plate,was added 2 μL of VRT stock solution in DMSO. The mixture was allowed toequilibrate for 10 mins at 30° C. The enzyme reaction was initiated bythe addition of 5 μL stock ATP solution prepared in assay buffer to afinal assay concentration of 240 μM. Initial rate data was determinedfrom the rate of change of absorbance at 340 nM (corresponding tostoichiometric consumption of NADH) using a Molecular Devices Spectramaxplate reader (Sunnyvale, Calif.) over 15 mins at 30° C. For each Kidetermination 12 data points covering the VRT concentration range of0-20 μM were obtained in duplicate (DMSO stocks were prepared from aninitial 10 mM VRT stock with subsequent 1:2 serial dilutions). Ki valueswere calculated from initial rate data by non-linear regression usingthe Prism software package (Prism 4.0a, Graphpad Software, San Diego,Calif.). Ki values are represented as A*<0.001 μM, A**<0.01 μM, A<0.05μM, B<0.5 μM, B*>0.7 μM, C*>1.25 μM, C**>2.0 μM, C<2.8 μM, D>2.8 μM,D*>4 μM.

A compounds are: 10, 16-18, 21-22, 24-25, 33-34, 37-39 and 43.

A* compounds are: 1, 2, 3-9, 11-15, 19-20, 23, 26-32, 40-42, and 44.

B compounds are 35, 36 and 45.

PKC Delta

An assay buffer solution was prepared which consisted of 100 mM HEPES(pH 7.5), 10 mM MgCl₂, 25 mM NaCl, 0.1 mM EDTA and 0.01% Brij. An enzymebuffer containing reagents to final assay concentrations of 0.002%Triton X-100, 200 μg/mL Phosphatidylserine, 20 μg/mL Diacylglycerol, 360μM NADH, 3 mM phosphoenolpyruvate, 70 μg/mL pyruvate kinase, 24 μg/mLlactate dehydrogenase, 2 mM DTT, 150 μM substrate peptide(ERMRPRKRQGSVRRRV SEQ ID NO. 2) and 46 nM PKC delta kinase was preparedin assay buffer. To 16 μL of this enzyme buffer, in a 384 well plate,was added 1 μL of VRT stock solution in DMSO. The mixture was allowed toequilibrate for 10 mins at 30° C. The enzyme reaction was initiated bythe addition of 16 μL stock ATP solution prepared in assay buffer to afinal assay concentration of 150 μM. Initial rate data was determinedfrom the rate of change of absorbance at 340 nM (corresponding tostoichiometric consumption of NADH) using a Molecular Devices Spectramaxplate reader (Sunnyvale, Calif.) over 15 mins at 30° C. For each Kidetermination 12 data points covering the VRT concentration range of0-20 μM were obtained in duplicate (DMSO stocks were prepared from aninitial 10 mM VRT stock with subsequent 1:2 serial dilutions). Ki valueswere calculated from initial rate data by non-linear regression usingthe Prism software package (Prism 4.0a, Graphpad Software, San Diego,Calif.).

A compounds are: 1, 5-9, 12, 15, 19-20, 23, 28, 29, 31, 40 and 42.

A** compounds are: 2, 11, 26, 27, 30, 32, and 44.

B compounds are: 3-4, 10, 13-14, 16-18, 21-22, 24, 25, 33, 37, 38, 39and 41.

C* compounds are: 34-36.43, and 45.

PKC Alpha

An assay buffer solution was prepared which consisted of 100 mM HEPES(pH 7.5), 10 mM MgCl₂, 25 mM NaCl, 0.1 mM EDTA, 100 μM CaCl₂ and 0.01%Brij. An enzyme buffer containing reagents to final assay concentrationsof 0.002% Triton X-100, 100 μg/mL Phosphatidylserine, 20 μg/mLDiacylglycerol, 360 μM NADH, 3 mM phosphoenolpyruvate, 70 μg/mL pyruvatekinase, 24 μg/mL lactate dehydrogenase, 2 mM DTT, 150 μM substratepeptide (RRRRRKGSFKRKA SEQ ID NO. 1) and 4.5 nM PKC alpha kinase wasprepared in assay buffer. To 16 μL of this enzyme buffer, in a 384 wellplate, was added 1 μL of VRT stock solution in DMSO. The mixture wasallowed to equilibrate for 10 mins at 30° C. The enzyme reaction wasinitiated by the addition of 16 μL stock ATP solution prepared in assaybuffer to a final assay concentration of 130 μM. Initial rate data wasdetermined from the rate of change of absorbance at 340 nM(corresponding to stoichiometric consumption of NADH) using a MolecularDevices Spectramax plate reader (Sunnyvale, Calif.) over 15 mins at 30°C. For each Ki determination 12 data points covering the VRTconcentration range of 0-20 μM were obtained in duplicate (DMSO stockswere prepared from an initial 10 mM VRT stock with subsequent 1:2 serialdilutions). Ki values were calculated from initial rate data bynon-linear regression using the Prism software package (Prism 4.0a,Graphpad Software, San Diego, Calif.).

B compounds are: 1, 2, 5, 7, 9, 12, 15, 20, 23, 26-30, 32, 40, 41, and44.

C compounds are: 3, 6, 8, 11, 13-14, 17, 19, 21-22, 31, 37, 38, and 42.

C* compounds are: 4, 10, 16, 18, 24, 25, 33-36, 39, 43 and 45.

While we have described a number of embodiments of this invention, it isapparent that our basic examples may be altered to provide otherembodiments that utilize the compounds, methods, and processes of thisinvention. Therefore, it will be appreciated that the scope of thisinvention is to be defined by the appended claims rather than by thespecific embodiments that have been represented by way of exampleherein.

We claim:
 1. A compound represented by the following structural formula:

or a pharmaceutically acceptable salt thereof, wherein: T is —NH— orabsent; each J_(c1) and J_(c2) is independently —CN, —F, —Cl, —OR,—CH₂OR, or —CF₃; each U₁, U₂, and U₃ is independently —H, Z, or J_(b)wherein no more than one of U₁, U₂, and U₃ is —H; or two of U₁, U₂, andU₃ join together to form a C1-C6 cyloalkyl ring having 0-1 heteroatomsindependently substituted with one or more J_(e); Z is Y2-Q2; Y2 isabsent or C1-6 alkyl optionally and independently substituted with oneor more J_(d); Q2 is absent or C3-C8 cyloalkyl having 0-1 heteroatomsoptionally and independently substituted with one or more J_(e), whereinY2 and Q2 are not both absent; each J_(b) is independently —F, —OR, —CN,—CF₃, —N(R)₂, —C(O)N(R)₂, C1-6 alkyl optionally and independentlysubstituted with one or more J_(a); each J_(a) is independently —F, —OR,—N(R)₂, or —C(O)N(R)₂; each J_(d) is independently —OR, —CN, —C(O)N(R)₂,—N(R)₂ or F; each J_(e) is independently C1-C6 alkyl, —OR, —N(R)₂, CF₃,or F; each R is —H or C1-C6 alkyl; and wherein there is a chiral centerat the carbon indicated by *, and wherein (S) represents stereochemistryon the ring carbon; with the proviso that the compound is not:


2. The compound of claim 1, wherein U₁ is Z and U₃ is J_(b).
 3. Thecompound of claim 2, wherein U₁ and U₂ are Z and U₃ is J_(b).
 4. Thecompound of claim 1, wherein Y2 is C1-C3 alkyl optionally andindependently substituted with one or more J_(d); Q2 is absent or C3-C6alkyl optionally and independently substituted with one or more J_(e);and each J_(d) is independently —OR, or F.
 5. The compound of claim 4,wherein J_(b) is —OH or —NH₂.
 6. The compound of claim 5, wherein J_(b)is —OH.
 7. The compound of claim 1, wherein each J_(c1) and J_(c2) isindependently —CF₃, —CN, —F, or —Cl.
 8. The compound of claim 7, whereineach J_(c1) and J_(c2) is independently —F, or —Cl.
 9. The compound ofclaim 8, wherein each J_(c1) and J_(c2) is —F.
 10. The compound of claim8, wherein J_(c1) is F and J_(c2) is Cl; or J_(c1) is Cl and J_(c2) isF.
 11. A compound represented by the following structural formula:

or a pharmaceutically acceptable salt thereof, wherein: T is —CH₂—,—CH(J_(b))-, —C(J_(b))₂-, —NH— or —N(J_(b))-; t is 0, 1, or 2; w is 0 or1; each J_(c) is independently CN, F, Cl, —OR, —CH₂OR, or CF₃; U is Z orJ_(b) Z is Y2-Q2; Y2 is absent or C1-6 alkyl optionally andindependently substituted with one or more J_(d); Q2 is absent or C3-C8cyloalkyl having 0-1 heteroatoms optionally and independentlysubstituted with one or more J_(e); wherein Y2 and Q2 are not bothabsent; each J_(b) is independently —F, —OR, —CN, —CF₃, —N(R)₂,—C(O)N(R)₂, C1-6 alkyl optionally and independently substituted with oneor more J_(a); each J_(a) is independently —F, —OR, —N(R)₂, or—C(O)N(R)₂; each J_(d) is independently —OR, —CN, —C(O)N(R)₂, —N(R)₂ orF; each J_(e) is independently —OR, CF₃, —N(R)₂ or F; and each R is —Hor C1-C6 alkyl; with the proviso that the compound is not:


12. A process for making a compound represented by structural formula I:

wherein: T is —NH— or absent; each J_(c1) and J_(c2) is independently—CN, —F, —Cl, —OR, —CH₂OR, or —CF₃; each U₁, U₂, and U₃ is independently—H, Z, or J_(b) wherein no more than one of U₁, U₂, and U₃ is —H; or twoof U₁, U₂, and U₃ join together to form a C1-C6 cyloalkyl ring having0-1 heteroatoms independently substituted with one or more J_(e); Z isY2-Q2; Y2 is absent or C1-6 alkyl optionally and independentlysubstituted with one or more J_(d); Q2 is absent or C3-C8 cyloalkylhaving 0-1 heteroatoms optionally and independently substituted with oneor more J_(e), wherein Y2 and Q2 are not both absent; each J_(b) isindependently —F, —OR, —CN, —CF₃, —N(R)₂, —C(O)N(R)₂, C1-6 alkyloptionally and independently substituted with one or more J_(a); eachJ_(a) is independently —F, —OR, —N(R)₂, or —C(O)N(R)₂; each J_(d) isindependently —OR, —CN, —C(O)N(R)₂, —N(R)₂ or F; each J_(e) isindependently C1-C6 alkyl, —OR, —N(R)₂CF₃, or F; each R is —H or C1-C6alkyl; and wherein there is a chiral center at the carbon indicated by*; and wherein (S) represents stereochemistry on the ring carbon, withthe proviso that the compound is not:

comprising the steps of: a) combining amide A with G to form C; b)heating C in the presence of hydrazine to form D; and c) displacing thehalogen on D with an amine J to form I.
 13. The process of claim 12,wherein step a) is conducted in the presence of lithium diisopropylamide(LDA).
 14. The process of claim 12, wherein the amine in step c) isprotected.
 15. The process of claim 12, wherein step c) is conducted inthe presence of suitable base, selected from the group comprisingpotassium carbonate, diisopropylethylamine (DIPEA), triethylamine, or1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), in a suitable solvent,selected from the group comprising dimethylformamide, dimethylsulfoxide(DMSO), or n-butanol (n-Bu-OH).
 16. The process of claim 12, whereinstep c) is conducted between 70° C. and 110° C.
 17. The process of claim12, wherein step c) is conducted using Pd as a catalyst.
 18. A processfor making a compound represented by structural formula I:

wherein: T is —NH— or absent; each J_(c1) and J_(c2) is independently—CN, —F, —Cl, —OR, —CH₂OR, or —CF₃; each U₁, U₂, and U₃ is independently—H, Z, or J_(b) wherein no more than one of U₁, U₂, and U₃ is —H; or twoof U₁, U₂, and U₃ join together to form a C1-C6 cyloalkyl ring having0-1 heteroatoms independently substituted with one or more J_(e); Z isY2-Q2; Y2 is absent or C1-6 alkyl optionally and independentlysubstituted with one or more J_(d); Q2 is absent or C3-C8 cyloalkylhaving 0-1 heteroatoms optionally and independently substituted with oneor more J_(e), wherein Y2 and Q2 are not both absent; each J_(b) isindependently —F, —OR, —CN, —CF₃, —N(R)₂, —C(O)N(R)₂, C1-6 alkyloptionally and independently substituted with one or more J_(a); eachJ_(a) is independently —F, —OR, —N(R)₂, or —C(O)N(R)₂; each J_(d) isindependently —OR, —CN, —C(O)N(R)₂, —N(R)₂ or F; each J_(e) isindependently C1-C6 alkyl, —OR, —N(R)₂ CF₃, or F; each R is —H or C1-C6alkyl; Pr is a protecting group; and wherein there is a chiral center atthe carbon indicated by *; and wherein (S) represents stereochemistry onthe ring carbon, with the proviso that the compound is not:

comprising the steps of: a) combining L with M to form N; and b)displacing the halogen on N with an amine J to form I.
 19. The processof claim 18, wherein L is combined with M in the presence of Pdcatalyst.
 20. The process of claim 18, wherein in step b) the amine isprotected.
 21. The process of claim 18, wherein step b) is conducted inthe presence of suitable base, selected from the group comprisingpotassium carbonate, diisopropylethylamine (DIPEA), triethylamine, or1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), in a suitable solvent,selected from the group comprising dimethylformamide, dimethylsulfoxide(DMSO), n-butanol (n-Bu-OH), or N-methylpyrrolidinone (NMP).
 22. Theprocess of claim 18, wherein step b) is conducted between 70° C. and110° C.
 23. The process of claim 18, wherein step b) is conducted usingPd as a catalyst.
 24. A process for making a compound represented bystructural formula I:

wherein: T is —NH— or absent; each J_(el) and J_(c2) is independently—CN, —F, —Cl, —OR, —CH₂OR, or —CF₃; each U₁, U₂, and U₃ is independently—H, Z, or J_(b) wherein no more than one of U₁, U₂, and U₃ is —H; or twoof U₁, U₂, and U₃ join together to form a C1-C6 cyloalkyl ring having0-1 heteroatoms independently substituted with one or more J_(e); Z isY2-Q2; Y2 is absent or C1-6 alkyl optionally and independentlysubstituted with one or more J_(d); Q2 is absent or C3-C8 cyloalkylhaving 0-1 heteroatoms optionally and independently substituted with oneor more J_(e), wherein Y2 and Q2 are not both absent; each J_(b) isindependently —F, —OR, —CN, —CF₃, —N(R)₂, —C(O)N(R)₂, C1-6 alkyloptionally and independently substituted with one or more J_(a); eachJ_(a) is independently —F, —OR, —N(R)₂, or —C(O)N(R)₂; each J_(d) isindependently —OR, —CN, —C(O)N(R)₂, —N(R)₂ or F; each J_(e) isindependently C1-C6 alkyl, —OR, —N(R)₂ CF₃, or F; each R is —H or C1-C6alkyl; Pr is a protecting group; and wherein there is a chiral center atthe carbon indicated by *; and wherein (S) represents stereochemistry onthe ring carbon, with the proviso that the compound is not:

comprising the steps of: a) combining J with M to form P; b) combining Pwith L to form Q; and c) deprotection of Q to form I.
 25. The process ofclaim 24, wherein step a) is conducted in the presence of a suitablebase selected from the group comprising potassium carbonate,diisopropylethylamine (DIPEA), triethylamine, or1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), in a suitable solvent,selected from the group comprising dimethylformamide, dimethylsulfoxide(DMSO), n-butanol (n-Bu-OH), or N-methylpyrrolidinone (NMP).
 26. Theprocess of claim 24, wherein step a) is conducted between 100° C. and130° C.
 27. The process of claim 24, wherein step b) is conducted in thepresence of Pd as a catalyst.
 28. A compound selected from thefollowing:

as well as diastereomers and enantiomers of compounds 4, 11, 13, 16, 20,23, 34, and 39, or a pharmaceutically acceptable salt thereof.
 29. Acomposition comprising a compound or pharmaceutically acceptable saltthereof according to any one of claim 1, 11, or 28, and apharmaceutically acceptable carrier, adjuvant, or vehicle.